研究小i机器人伴侣

1一、引言2007年,生物大分子国家重点实验室各项工作成绩斐然.
实验室新增各类科研项目30项,其中:科技部项目6项,国家基金委项目14项,中科院知识创新工程重要方向项目10项.
在SCI收录的刊物上发表论文94篇,其中IF〉10的论文12篇,篇均IF=4.
72.
申请国内发明专利12项,国外发明专利2项,有2项专利被授权.
梁伟研究员"肿瘤靶向的新型纳米药物输送系统"成果入选2007年中国十大科技进展.
实验室始终坚持人才是第一资源的理念,在各级领导和主管部门的大力支持下,引进了一批杰出的中青年科学家.
2007年引进"百人计划"2名,实验室目前已初步建成一支由国际著名或知名科学家领衔的,老中青相结合,规模适度、结构合理、充满活力的高素质创新科技队伍.
实验室现由35名研究员组成(其中院士7名;中国科学院"百人计划"入选者19名;国家杰出青年基金获得者5名;"长江学者奖励计划"特聘教授2名).
在读研究生301名,其中135名博士生,162名硕士生.
获得博士学位27名,24名博士后.
2007年,经过各方积极不懈的努力,生物物理所蛋白质与分子生物医学科研楼正式破土动工,预计将于2008年中完成土建工程,2008年底投入使用,这将大大改善实验室的科研用房条件.
2007年末,中科院第12次院长办公会审议并原则通过了中国科学院蛋白质科学平台二期建设实施方案,这是继生物物理所于2005年底完成了研究平台的一期建设后,科学院大力支持科研条件建设的又一重大举措,平台二期建设是一期建设的跨越式提升,通过持续的平台建设,最终将建成大规模、高产出、国际一流的蛋白质科学研究平台,为提升我国生命科学的自主创新能力、不断产出国际水平的原始创新成果、保障经济社会可持续发展做出历史性贡献.

科技部批准以生物大分子国家重点实验室为基础建设"蛋白质科学国家实验室".
目前,在所领导和全室同仁共同努力和积极推进下,蛋白质科学国家实验室的筹备工作已全面启动.

先进的价值理念,良好的文化生态,和谐的科研氛围,已经成为现代实验室核心竞争力的重要源泉.
展望2008年,将是实验室发展至关重要的一年.
我们将积极配合研究所推进科研条件建设,着力打造蛋白质研究国家基地;完善技术支撑体系建设,以技术创新推动科学创新;加强学科方向的研讨,凝练出若干世界科学难题,组织力量开展攻关,力争为我国生命科学发展作出历史性贡献;加强国际合作与交流,着力推进与国际一流研究机构建立长期稳定的战略合作伙伴关系,不断提升实验室在国际同行中的学术地位.
生物大分子国家重点实验室主任:生物大分子国家重点实验室学委会主任:2一、IntroductionIn2007theNationalKeyLaboratoryofBiomacromoleculeshadmanyexcellentacademicachievements.
NotonlyhasourLabpublishedhighqualitypapers,butthestandardofscientificresearchinourLabhasimprovedcontinuously.
NinetyfourpaperswerepublishedinSCIjournals,ofwhich12hadimpactfactorsgreaterthan10.
Theaverageimpactfactorperpaperwas4.
72.
TheLabappliedfor11domesticpatentsandtwooverseaspatents,twoofwhichweregranted.
ProfessorWeiLiang's,'NewNanoDrugDeliverySystemforTargetingTumors'wasselectedasoneofthe'TenOutstandingScienceandTechnologyAchievementsofChina'in2007.
Inrecentyears,thecompetitivestrengthandoverallabilityoftheLabtoundertakeimportantnationalresearchprogramshasimprovedremarkably.
In2007,theLabundertookabout30newscienceresearchprojects,including6projectsfromtheMinistryofScienceandTechnology,14projectsfromtheNationalNaturalScienceFoundationofChina,and10projectsfromtheKnowledgeInnovationProgramoftheChineseAcademicofSciences.
TheMinistryofScienceandTechnologyauthorizedIBPtoestablishtheNationalLaboratoryofProteinScienceonthefoundationslaidbytheNationalKeyLaboratoryofBiomacromolecules.
ThankstotheeffortsoftheDirectorsofIBPandthemembersoftheLab,preparationsfortheestablishmentoftheNationalLabofProteinSciencearealreadyunderway.
TheNationalKeyLaboratoryofBiomacromoleculesinsiststhattalentisitsgreatestresource.
WiththestrongsupportoftheDirectorsandadministrationstaff,theLabhasattractedagroupofoutstandingyoungandmiddle-agedscientists.
In2007,theLabrecruitedtwoscientistsinthe'Onehundredtalentsprogram'.
OurLabhasagood-sized,well-structuredteamoftop-levelyoung,middle-agedandolderscientificresearchersledbyinternationallyfamousscientists.
Thereare36professors,ofwhichsevenaremembersoftheCAS,19aremembersofCAS's'Onehundredtalentsprogram',fivearewinnersof'NationalOutstandingYouthFoundation'grants,andtwohaveprofessorialchairsaspartofthe'ChangJiangScholarsProgram'.
Thereare301graduatestudentsintheLab,ofwhich135arestudentspursuingPhDdegrees,and162studentsarepursuingMaster'sdegrees.
Twenty-sevenresearchersintheLabhavePhDdegrees,and24arecurrentlydoingpostdoctoralresearch.
In2007,thankstounrelentingeffortsoneachside,IBPstartedtobuildtheProteinandMolecularMedicalSciencesResearchBuilding.
Itisexpectedtobefinishedinthemiddleof2008,andwillbeinusebytheendof2008.
Thiswillgreatlyimproveourscientificresearchfacilities.
ThesecondphaseoftheconstructionandimplementationplanfortheCASproteinsciencesplatformwasdiscussedandapprovedbytheCASPresident'sOfficeattheendof2007.
ThisrepresentsafurtherinvestmentbyCAStoimprovethescientificresearchenvironmentatIBPfollowingonfromtheestablishmentoftheproteinscienceplatformattheendof2005.
Thesecondphaseofconstructionwilldeliverfurtherimprovementsonthefoundationestablishedbythefirstphase,givingrisetoalargescale,highoutputandinternationallytoplevelproteinscienceresearchfacilitywhichwillmakeanhistoriccontributiontoimprovingtheinnovativeabilityofnationallifesciences,producehighlevelandoriginal3innovativescientificachievementsandcontributetothesustainabledevelopmentoftheeconomy.
Advancedvalues,ahealthyworkculture,andaharmoniousscientificresearchenvironmenthavealreadybecomeanimportantsourceofourLab'scorecompetitiveability.
2008willbeaveryimportantyearinthedevelopmentoftheLaboratory.
IncooperationwiththeInstitutetoimprovescientificresearchconditions,wewillbuildanationalresearchbaseforproteinscience,improvethetechnologysupportsystem,promotescientificinnovationthroughtechnologyinnovation,strengthenconsultationoverresearchdirections,organizescientiststounravelanumberoftheworld'sdifficultscienceproblems,andmakeanhistoriccontributiontothedevelopmentofthelifesciencesinChina.
TheLabwillaimtopromoteinternationalacademicexchangesandcollaborations,andestablishsteadyandlongtermrelationshipswithadvancedinternationalscientificresearchinstitutes,sothattheLabwillcontinuallyimproveitshighacademicreputationamongotherinternationalscientificresearchinstitutesinitsfield.
NationalLaboratoryofBiomacromeleculesDirector:TaoXuAcademicCouncilChairman:Chih-chenWang4二、代表性成果(SelectedAchievements)1、T细胞控制天然免疫细胞产生的炎症反应Adaptiveimmunecellstemperinitialinnateresponses.
NATUREMEDICINE2007,13(10):1248-1252唐宏研究组和付阳心研究组发现在肝炎病毒感染的极早期(<2天),未被激活的T细胞对于控制天然免疫细胞产生的炎症反应期至关重要的抑制作用.
病毒感染适应性免疫系统缺陷的裸鼠或剔除T细胞的小鼠后,小鼠因天然免疫系统被激活导致的炎症因子飙升而剧烈死亡,过继性输入T细胞或者进一步剔除自然杀伤细胞(NK)后小鼠重新存活,炎症反应也得到了有效抑制.
研究成果加深了人们对于炎症反应的认识,并提出了T细胞参与天然免疫反应的负性调控的新理论.
对于临床上深入了解病毒性感染的炎症反应和病毒清除机理,免疫低下病人(新生儿,老年人,器官移植患者或艾滋病人)机会性感染的控制具有极高的理论价值.
ThejointresearchbyDrs.
HongTangandYang-xinFu'slabshasshownthatTcellsoftheadaptiveimmunesystemsuppressoverzealousearlyinnateresponses-"cytokinestorm"-toinfectionthatusuallyleadtosevereimmunopathologyandhighdeathrates.
Theyhavebeenabletoshow,throughthedepletionofCD4+andCD8+Tcellsinwild-typemiceorbyadoptivetransferofTlymphocytesintoRag-1deficientmice(lackofTcells),thatTcellsarebothnecessaryandsufficienttotempertheearlyinnateresponse.
Theyfurtherdemonstratedthatviralinfectionoradministrationofasynthetic,RNAvirusgenomemimickerycompound,poly(I:C),ledtocytokinestorminT-cell-deficientmiceinNKcellsandtumornecrosisfactor(TNF)dependentmanner.
Theyalsofoundthat,inadditiontotheconventionalinhibitoryTcells(Tregcells),closecontactofrestingnon-Treg(CD4+CD25–Foxp3–)orCD8+Tcellswithinnatecellscouldalsosuppressthecytokinesurgeinanantigen-independentfashion.
Thesefindingsthereforesuggestthatearlyinnateimmunityrequirestheadaptiveimmunityinchecktomaintaintheappropriateimmuneresponses.
Thesefindingsmayalsoprovideanovelmechanismofimmunopathologyduringacuteinfectionsandpotentialanti-inflammationregime.
2、UNC-31蛋白调控线虫致密核心囊泡锚定的机理PKAActivationBypassestheRequirementforUNC-31intheDockingofDenseCoreVesiclesfromC.
elegansNeurons.
Neuron2007,56:657–669线虫是很好的研究遗传和发育的系统,但其在细胞生物学特别是囊泡转运与分泌领域的贡献却5十分有限,其主要原因是缺少高时空分辨的研究手段.
2007年11月21日,《Neuron》发表了徐涛研究组在该领域的最新成果.
他们克服了研究手段上局限,发展了模式生物线虫的单细胞分离和培养方法,首次在线虫单神经元上用膜电容检测技术记录到胞吐和胞吞过程,结合改进的碳纤微电极技术和囊泡转运的显微成像技术等先进的生物物理方法,将高时空分辨的分泌检测技术应用在线虫上,建立了在线虫细胞水平研究调控型分泌的技术平台.
利用该技术平台,证明了核心致密囊泡的胞吐过程需要一种称为UNC-31(CAPS在线虫中的同源蛋白)的蛋白,阐明了该蛋白参与囊泡锚定的作用机制,并发现了UNC-13(Munc13-1在线虫中的同源蛋白)与UNC-31蛋白之间存在相互作用.
该工作开辟了利用线虫模式生物研究囊泡分泌的新方向.
ThenematodeC.
elegansprovidesapowerfulmodelsystemforexploringthemolecularbasisofsynaptogenesisandneurotransmission.
However,thelackofdirectfunctionalassaysofreleaseprocesseshaslargelypreventedanindepthunderstandingofthemechanismofvesicularexocytosisandendocytosisinC.
elegans.
Forthefirsttime,weestablishahighspatial-temporalmethodtomonitorsecretionfromneurons.
Wedevelopeddirectelectrophysiologicalassays,includingmembranecapacitanceandamperometrymeasurements,inprimaryculturedC.
elegansneurons.
Inaddition,wehavesucceededinmonitoringthedockingandfusionofsingledensecorevesicles(DCVs)employingtotalinternalreflectionfluorescencemicroscopy.
Withtheseapproachesandmutantperturbationanalysis,weprovidedirectevidencethatUNC-31isrequiredforthedockingofDCVsattheplasmamembrane.
Interestingly,thedefectinDCVdockingcausedbyUNC-31mutationcanbefullyrescuedbyPKAactivation.
WealsodemonstratethatUNC-31isrequiredforUNC-13-mediatedaugmentationofDCVexocytosis.
Thisworkrepresentsbothasignificanttechnicaladvanceintheanalysisofregulatedexocytosisinanimportantgeneticsystem,C.
elegans,butalsouncoversnewinsightsintothemechanismsunderlyingsecretion.
3、脂肪细胞GLUT4贮存囊泡GSV的锚定DissectingmultiplestepsofGLUT4traffickingandidentifyingthesitesofinsulinaction.
CELLMETAB2007,5:47-57徐涛研究组开发了一种分辨脂肪细胞GLUT4贮存囊泡GSV与细胞质膜融合过程中锚定、启动、融合等关键调控步骤的方法,发现了胰岛素在提高GSV囊泡锚定速率的同时,更关键的调控步骤是在囊泡锚定在细胞膜下之后使囊泡具有融合能力的启动过程.
研究还发现胰岛素激活的PI3K及其下游效应物AS160调控了囊泡的锚定过程.
研究组同时利用全内反射荧光显微成像方法发现和研究GLUT4贮存6囊泡的调控作用,挑战了之前的GLUT4转运调控过程,为研究胰岛素信号途径提供了重要资料.
2007年1月《DevelopmentalCell》发表的评论文章认为:该研究方法有利于解决胰岛素怎样调控GSV囊泡与质膜融合等重要科学问题,有利于将胰岛素对GSV调控机制的研究重点转向囊泡与质膜融合这一关键步骤.
Insulin-stimulatedGLUT4translocationiscentraltoglucosehomeostasis.
FunctionalassaystodistinguishindividualstepsintheGLUT4translocationprocessarelacking,thuslimitingprogresstowardelucidationoftheunderlyingmolecularmechanism.
Firstwehavedevelopedarobustmethod,whichreliesondynamictrackingofsingleGLUT4vesiclesinrealtime,fordissectingandsystematicallyanalyzingthedocking,priming,andfusionstepsofGLUT4vesicleswiththecellsurfaceinlivingadipocytes.
Usingthismethod,wehaveshownthatthepreparationofGLUT4vesiclesforfusioncompetenceafterdockingatthesurfaceisakeystepregulatedbyinsulin,whereasthedockingstepisregulatedbyPI3Kanditsdownstreameffector,theRabGAPAS160.
ThesedatashowthatAkt-dependentphosphorylationofAS160isnotthemajorregulatedstepinGLUT4trafficking,implicatingalternativeAktsubstratesoralternativesignalingpathwaysdownstreamofvesicledockingatthecellsurfaceasthemajorregulatorynode.
4、PEG-PE聚合物胶束与阿霉素的自组装过程与其性质Improvingpenetrationintumorswithnanoassembliesofphospholipidsanddoxorubicin.
JNATCANCERINST.
2007,99(4):1004-1015梁伟研究组的工作表明:聚乙二醇衍生化磷脂与抗肿瘤化疗药物-阿霉素可自组装形成纳米尺度的新型输送载体,提高阿霉素在肿瘤组织中的富集和对深层组织细胞的渗透,进而增强了阿霉素的抗肿瘤效果并降低了毒性.
研究首次证明了包载阿霉素的聚乙二醇衍生化磷脂纳米胶束可以选择性地在肿瘤组织蓄积并渗透到深层肿瘤组织提高肿瘤细胞内药物浓度,从而显著增强阿霉素的细胞毒性、抑制肿瘤的生长、延长小鼠的生存时间和降低药物的毒性,为临床治疗肿瘤提供了新的有效手段.
该技术已申请国际专利,具有自主知识产权.
同期杂志配发评论指出,这是一个简单但能有效地将药物和合适的载体整合起来产生很好效果的例子,也许药理学概念上的抗肿瘤药物由于不能正确识别它们的靶标而错伤病人的时代将会结束.
ThestudybyWeiLiang'sgrouphasindicatethatPEG-PEanddoxorubicinself-assembletoformanovelnano-carrier,whichimprovestheantitumoractivityofdoxorubicinandreducesitstoxicicitybyenhancingtheaccumulationandpenetrationofdoxorubicininthecellslocatedinthedeep-layersofthetumor.
Thisisfirsttimetoprovethatthedoxorubicin-loadedPEG-PEmicellesselectivelyaccumulateandpenetrateinthedeepertumortissuetoincreasetheintracellulardrugconcentrationintumor.
Thus,doxorubicinloaded7PEG-PEmicellesgreatlyenhancesthecytotoxicityofdoxorubicin,inhibitsthetumorproliferation,prolongsthesurvivaltimeoftumor-bearmiceandreducesthedrugsystemictoxicity.
Thisdrugpackagingtechnologymayprovideanewstrategyfordesigncancertherapy.
Meanwhile,thistechnologyhasbeenappliedforinternationalpatentwithourownintellectualpropertyrights.
TheeditorialintheJNatlCancerInsthaspointedout:thisstudyisasimplebuteffectivedemonstrationofthebenefitsofintegrationofadrugwithanappropriatecarriertoyieldastrikinggaininefficacy.
Maythedaysofpharmacologicmissilesthatmisstheirtargetandfriendlyfirethatkillspatientssoonbeover.
5、人源多功能转录共激活因子p100的三维精细结构Themultifunctionalhumanp100protein'hooks'methylatedligands.
NatStructMolBiol2007,14(8):779-784刘志杰课题组报告了人源p100这种多功能转录共激活因子的三维精细结构,解释了多功能转录共激活因子p100在转录和剪接中的不同作用,有利于进一步了解p100蛋白的功能和作用机制.
表明多功能转录共激活因子p100及与其特异性结合的多蛋白复合体在人体免疫反应的IL-4信号传导通路中起着非常重要的转录、调控和激活作用.
研究还发现多功能转录共激活因子p100的一种全新的钩状结构域分布方式并将其命名为TSN结构域.
Thehumanp100proteinisavitalregulatorofcellulartranscriptionprocesses.
p100hasbeenshowntophysicallybridgepromoterspecificactivatorswiththebasaltranscriptionmachineryresultinginanincreaseinthelevelofgenetranscription.
Here,wedemonstrateinteractionbetweenthetudordomainofp100andtheUsnRNPcomplex,thussuggestingaroleforp100intheprocessingofpre-mRNA.
Wedeterminedthecrystalstructureofp100SN-Tudordomaininordertodelineatethemolecularbasisoftheproposedfunctionsforp100.
Theinterdigitatedstructureresemblesahook,withahingecontrollingthemovementandorientationofthehook.
OurstudiessuggestaconservedaromaticcagehooksmethylgroupsofsnRNAsandanchorsthep100tothespliceosome.
Thefindingsofthisstudyprovideimportantstructuralinsightsthatpartlyexplainthedistinctrolesofp100intranscriptionandsplicing.
6、模拟流感演化的计算机新模型的建立Evolution.
GENOMERESEARCH2007,了解流感病毒的演化规律对防治流感起着非常重要的意义.
尽管流感基因组看似简单,但流感变化分子规律却难以捕捉.
蒋太交研究员实验室首次提出了利用网络模型来描述共进化的基因组信HINGEHOOKHINGEHOOK8息,创造性地将每个病毒表示为一个基因组元件相关性网络.
该研究工作还引进几个网络特性参数(连接度K、连接度变化R以及表征片断间协同进化程度的量C),定量地描述出流感进化中的很多重要特征:抗原演化及其结构基础,流感片断间的功能联系及片断间的重组.
该文章的三个评审专家都认为这个新方法将推动流感演化的分子机制的了解.
同时他们认为该新颍的网络模型是研究流感病毒全基因组演化的重要方法,代表了该领域的重要进展.
UnderstandingtheevolutionofinfluenzaAvirus,whichposesaglobalchallengetopublichealth,isofspecialsignificanceforitscontrolandprevention.
Althoughthegenomestructureofthevirusisseeminglysimple,theirevolutionarypatternsandmolecularmechanismsaredifficulttoreveal.
TherecentavailabilityoffullgenomicsequencedataforalargenumberofhumaninfluenzaA(H3N2)virusisolatesovermanyyearshaveprovidedanopportunitytoanalyzeitsevolutionbyconsideringallgenesegmentssimultaneously.
However,suchanalysisrequiresnewcomputationalmodelsthatcancapturethecomplexevolutionaryfeaturesovertheentiregenome.
AsbeingreportedonlinebyGenomeResearchon21November,aresearchgroupheadedbyProf.
JiangTaijiaowiththeCASInstituteofBiophysicshassetupanewcomputationalmodeltodepictmanyepidemiologicalcharacteristicsoftheA(H3N2)virus.
Byanalyzingtheco-occurrenceofthenucleotidesintheentiregenomeofthevirus,Prof.
Jiangandcolleagueshavedevelopedanetworkmodeltodescribeitsevolutionarypatternsanddynamics.
Thenetworkmodelcaneffectivelyidentifytheantigenicfeaturesofthevirusatthewhole-genomelevelandaccuratelydistinguishthecomplexpatternsintheviralevolutionbetweendifferentgenesegments.
Theirworkfurthershowsthattheco-occurringnucleotidemodulesapparentlyunderpinthedynamicsoftheH3N2viralevolution.
Moreover,thenetworkmodelallowedthemtoidentifykeyaminoacidsubstitutionsthatdictatetheantigenicevolutionofhumanH3N2virus.
Thestudydemonstratesthatthenetworkofnucleotideco-occurrencespresentsapromisingmethodfortrackingdowntherouteoftheviralevolution.
Theideathatanetworkcanmodelitsevolutionisastreamlinedapproachtofurtherunderstandandpredictthepatternsofglobalepidemicsandwillbeagreatstepforwardinvirusbiology,"accordingtoareviewerofthepaper.
Thisworkisalsohighlyappreciatedbyanotherreviewer,"Fewpapersonnovelmethodsfortheanalysisoflargecollectionsofinfluenzagenomeshavebeenpublishedtodate,andthispaperrepresentsasignificantadvance.
97、ZAP通过招募RNA外切酶加工复合体降解靶RNAThezinc-fingerantiviralproteinrecruitstheRNAprocessingexosometodegradethetargetmRNA.
PNAS2007,104(1):151-156宿主抗病毒因子锌指结构抗病毒蛋白(ZAP)能够通过防止病毒mRNA在细胞质中的积累从而特异性抑制小鼠白血病病毒和辛德比斯病毒的复制.
高光侠课题组以前的工作表明,ZAP直接结合特异性的靶mRNA.
本文中,提供一些证据表明ZAP招募RNA外切酶加工复合体(Exosome)降解靶RNA:在蔗糖或甘油梯度离心中ZAP与Exosome共迁移;ZAP的免疫沉淀可以共沉淀下来Exosome组分;体外结合实验结果表明ZAP直接与Exosome组分hRrp46p相结合,ZAP与之结合的功能域定位在224-254氨基酸;利用RNAi方法将Exosome组分hRrp46p或hRrp41p敲低后ZAP的活性被减弱.
上述结果表明ZAP是一种调控mRNA稳定性的反式作用因子.
Thezincfingerantiviralprotein(ZAP)isahostantiviralfactorwhichspecificallyinhibitsthereplicationofMoloneymurineleukemiavirusandSindbisvirusbypreventingaccumulationoftheviralmRNAinthecytoplasm.
Inpreviousstudies,wedemonstratedthatZAPdirectlybindstoitsspecifictargetmRNAs.
Inthisreport,weprovideevidenceindicatingthatZAPrecruitstheRNAprocessingexosometodegradethetargetRNA.
ZAPco-migratedwiththeexosomeinsucroseorglycerolvelocitygradientcentrifugation.
ImmunoprecipitationofZAPcoprecipitatedtheexosomecomponents.
Invitropull-downassaysindicatedthatZAPdirectlyinteractedwiththeexosomecomponenthRrp46pandthatthebindingregionofZAPwasmappedtoaminoacids224-254.
DepletionoftheexosomecomponenthRrp41porhRrp46pwithsmallinterferingRNAsignificantlyreducedZAP'sdestabilizingactivity.
ThesefindingssuggestthatZAPisatypeoftrans-actingfactormodulatingmRNAstability.
8、氧化铁纳米颗粒具有类似过氧化物酶的催化活性Intrinsicperoxidase-likeactivityofferromagneticnanoparticles.
NATURENANO2007,2:577-583阎锡蕴研究组研究发现氧化铁纳米颗粒具有过氧化物酶的催化功能.
研究组还利用纳米颗粒模拟酶的这一新特性,设计了多种免疫检测方法,实现了对乙肝病毒表面抗原和肌钙蛋白的检测.
同期杂志上还刊登了中佛罗里达大学(UniversityofCentralFlorida)纳米科学技术中10心J.
ManuelPerez教授写的述评:《氧化铁纳米颗粒:蕴藏的功能》.
该述评称阎锡蕴、柯莎及其课题组成员首次发现氧化铁纳米颗粒具有类似过氧化物酶的催化活性,并提出了氧化铁纳米颗粒模拟酶的概念.
虽然如何在生物技术和医疗领域更好地利用纳米材料的催化活性还有待探索,但氧化铁纳米颗粒催化活性的发现将使人们对此产生更多的关注.
Prof.
XiyunYan'sgroupreportedthatmagnetitenanoparticlespossessanintrinsicenzymemimeticactivitysimilartonaturalperoxidases.
Basedonthenovelfindings,theydevelopedanovelimmunoassayusingthemulti-functionofantibody-modifiednanoparticles(targeting,separationandcatalysis).
AcommentaryarticleinthesameissueofNatureNanotechnologynotedthatYan,Perrettandcolleagues'resultsshowforthefirsttimethatironoxidenanoparticlescanfunctionasperoxidasenanomimetics.
Althoughthepotentialwideapplicationsoftheperoxidase-likeactivityofironoxidenanoparticlesinmedicineandbiotechnologyremaintobeexplored,the'ready-made'simplicityofcatalyticactivityinironoxidenanoparticleswillcertainlyattractalotofattention.
11三、组织机构(Organization)(一)室务委员会实验室主任:徐涛研究员实验室副主任:龚为民研究员阎锡蕴研究员唐宏研究员(二)学术委员会学术委员会主任:王志珍院士学术委员会成员:(按姓氏拼音排序)姓名职称工作单位常文瑞院士中科院生物物理所郭爱克院士上海生科院神经所/中科院生物物理所贺福初院士军事医学科学院蒋华良研究员中科院上海生科院药物所李家洋院士中国科学院李林研究员中科院上海生科院生化所林其谁院士中科院上海生科院生化所强伯勤院士中国医学科学院秦志海研究员中科院生物物理所饶子和院士南开大学/中科院生物物理所施蕴渝院士中国科技大学田志刚研究员中国科技大学王大成院士中科院生物物理所徐涛研究员中科院生物物理所于军研究员中科院基因组所张先恩研究员科技部基础司(三)实验室成员(按姓氏拼音排序)毕利军常文瑞陈畅陈润生邓红雨范祖森高光侠龚为民杭海英姬广聚江涛蒋太交焦仁杰柯莎梁栋材梁伟刘迎芳刘志杰马跃苗龙秦燕秦志海饶子和孙飞唐宏唐捷王大成王江云王盛典王志珍王志新徐涛阎锡蕴杨福愉殷勤伟12四、管理(Management)(一)实验室管理实验室依据"国家重点实验室建设与管理暂行办法"对实验室进行全面管理.
本室实行实验室主任负责制、学术委员会评审制;实验室采用以研究方向为导向,以研究组为基本科研单元的运行模式,每年2-3月间召开上年度学术年会.
实验室主任就年度运行管理、经费使用提出可行方案,并向学术委员会汇报上年度的实验室工作总结.
(二)学术委员会学术委员会结合本室学术年会每年召开全体会议一次,评议实验室的工作,内容包括确定实验室研究方向、制定及修改课题指南、审批课题申请、检查课题进展情况、监督经费使用、评审科研成果及审议学术活动计划等.
(三)研究方向1.
蛋白质三维结构与功能研究2.
蛋白质功能与折叠原理研究3.
生物膜和膜蛋白功能结构研究4.
计算与系统生物学5.
感染与免疫的分子基础6.
蛋白质药物与多肽药物(四)课题管理实验室成员每年应按时向实验室秘书提交以下材料:1)当年发表的具有"生物大分子国家重点实验室"署名的全部著作目录(包括专著、论文,国际及全国性学术会议论文等),并提交版面清楚平整的论文单印本一式一份及论文电子版;2)当年获得国际、国家或省部级科技奖励的证书复印件一份;3)年度工作报告(中、英文各一份)的电子文档.
报告格式按实验室秘书提供的文档模版填写.
13五、研究进展(ResearchProgressReport)(一)蛋白质三维结构与功能研究1、常文瑞组在光合膜蛋白研究方面,我们完成了黄瓜LHCII晶体结构的测定工作,发现LHCII分子中两个Lutein分子的构象差异,提出这种构象改变可能与光淬灭位点的形成有关;发现不同存放期的LHCII晶体的晶胞参数和光谱特征有所改变,推测这种变化可能与其分子构象变化有关,并将可能为研究其光能传递和光保护提供新的信息,部分研究结果已经发表(Biochem.

Biophys.
Res.
Commun.
),后续研究工作正在进展之中.
另外我们已成功获得高等植物光系统II中几种光合膜蛋白及其复合物的样品,其中数种样品已获微晶,晶体质量正在改善和优化之中.

在可溶性的重要功能蛋白研究方面,我们成功地用基于Br的SAD法测定了马来酰丙酮酸异构酶MPI高分辨率晶体结构,这是第一个MSH依赖性酶的晶体结构(J.
Biol.
Chem.
).
并解析了人源S100A13蛋白的晶体结构,发现一些独特的结构特征(Biochem.
Biophys.
Res.
Commun.
).
解析了人胞质磺酰基转移酶SULT1A2与PAP复合物的晶体结构,研究结果已经投送国际核心杂志.
解析了硫氧化还原酶(SOR)的两种不同晶型的晶体结构,研究结果正在总结之中.
此外还分别测定了蚯蚓纤溶酶三种组分:EFE-b、EFE-c、EFE-g和牛胰蛋白酶抑制剂aprotinin的复合物的晶体结构.
并获得人源DPY-30的晶体,正在进行重原子衍生物的制备工作.
1、Prof.
ChangW.
R.
WehavesolvedLHC-IIcrystalstructurefromcucumberat2.
66resolution.
Itwasfoundthattwoluteinmoleculeshavedifferentconformations.
Weproposedthattheconformationalchangemightrelatetoformationofnon-photochemicalquenchingsitebasedonthestructureanalysis.
WealsodiscoveredthatthedehydrationoftheLHCIIcrystalsresultedinanotableshrinkageofthecrystalunitcelldimensions,whichwasaccompaniedbyared-shiftofthefluorescenceemissionspectraofthecrystals.
ThesephenomenasuggestthechangesinthecrystalpackingduringdehydrationmightbethecauseofinternalconformationalchangeswithinLHCII.
Partoftheresultswaspublished(Biochem.
Biophys.
Res.
Commun.
).
Moreover,wehavesuccessfullyharvestedsomeamountofseveralmembraneproteinsandcomplexesfromhigherplantphotosystemII,andgainmicrocrystalsofsomesamples,nowwearetryingtooptimizethecrystalgrowthconditions.
Ontheotherprojects,wehavesolvedhighresolutionstructuresofMPI,whichisthefirstexampleofMSH-dependentenzyme,usingSADmethod(J.
Biol.
Chem.
).
AndwehavesolvedcrystalstructureofhumanS100A13andfoundsomeuniquestructuralfeatures(Biochem.
Biophys.
Res.
Commun.
).
Inaddition,crystalstructureofSULT1A2complexedwithPAP,crystalstructuresofsulfuroxygenasereductase(SOR)intwocrystalforms,threestructuresofEFEcomponentscomplexedwithinhibitorwerealsosolved,theresultsweresubmittedorinpreparation.
AndwehavesuccessfullycrystallizedhumanDPY-30,thescreeningofheavyatomderivativesareinprogress.
发表文章:1、WangR,YinYJ,WangF,LiM,FengJ,ZhangHM,ZhangJP,LiuSJ,ChangWR.
Crystalstructuresandsite-directedmutagenesisofamycothiol-dependentenzymerevealanovelfoldingandmolecularbasisformycothiol-mediatedmaleylpyruvateisomerization.
JBiolChem.
2007;82(22):16288-294.
2、LiM,ZhangPF,PanXW,ChangWR.
CrystalstructurestudyonhumanS100A13at2.
0Aresolution.
BiochemBiophysResCommun.
2007;356(3):616-21.
3、YanH,ZhangP,WangC,LiuZ,ChangWR.
TwoluteinmoleculesinLHCIIhavedifferentconformationsandfunctions:Insightsintothemolecularmechanismofthermaldissipationinplants.
BochemBiophysResCommun.
2007;355(2):457-63.
142、龚为民组目前主要集中在蛋白质翻译起始、生物膜转运等相关的蛋白质复合体和膜蛋白的结构生物学研究,建立了膜蛋白和蛋白质复合体的表达纯化体系,形成了相关功能研究的稳定合作伙伴.
在真核翻译起始复合物及其与核糖体相互作用方面取得重要进展,共解析其中四个真核翻译起始因子亚基的晶体结构,表达纯化了eIF3和mini-MFC两中复合体,纯化了酵母核糖体并完成了亚基拆分,为后续解析起始复合体的三维结构和研究起始复合体与核糖体的相互作用打下了基础.
根据生物膜转运的功能研究,筛选并大量可溶表达出22种膜转运相关的人源未知结构蛋白,完成了TRAPP复合体中synbindin蛋白的晶体结构和NMR结构,研究了其与syndecan的相互作用.
表达纯化出三种人源膜蛋白,其中一种已获得微晶.
另外我们还完成了一些酶催化机制的结构生物学研究.

2、Prof.
GongW.
M.
Ourresearchinterestsarethestructuralbiologystudiesofproteinsandproteincomplexesinvolvedintranslationinitiation,vesicletrafficking.
Wehavesetupsomeexpressionandpurificationsystemsformembraneproteinsandproteincomplexesandestablishedstablecollaborationswithothergroupsonrelatedfunctionalstudies.
Wehavealreadymadesignificantprogressonthesamplepreparationofeukaryotictranslationinitiationcomplexesandyeastribosome.
Fourcrystalstructuresofeukaryoticinitiationfactorsubunitshavebeensolvedinourlab.
Basedonthefunctionalstudies,weselectedandpurified22humanvesicletraffickingrelatedproteinswithunknownstructuresinsolubleform.
Thestructureofsynbindin,acomponentofTRAPPcomplex,hasbeensolvedbycrystallographyandNMR.
Wealsohaveexpressedandpurifiedthreehumanmembraneproteins,oneofwhichhasbeencrystallizedasamicro-crystal.
Besides,wehavefinishedotherstructuralbiologystudiesonenzymecatalyticmechanisms.
发表文章:1.
HouX,LiuR,RossS,SmartEJ,ZhuH,GongW.
CrystallographicStudiesofHumanMitoNEET.
JBiolChem.
2007;282(46):33242-6.
2.
JLi,CZhang,XXu,JWang,HYu,RLai,WGong.
Trypsininhibitoryloopisanexcellentleadstructuretodesignserineproteaseinhibitorsandantimicrobialpeptides.
FASEBJ.
2007;21(10):2466-73.
3.
WangM,LiuL,WangY,WeiZ,ZhangP,LiY,JiangX,XuH,WGong.
CrystalstructureofhomoserineO-acetyltransferasefromLeptospirainterrogans.
Biochem.
Bioph.
Res.
Co.
2007;363(4):1050-6.
4.
XHou,YWang,ZZhou,SBao,YLin,WGong.
CrystalstructureofSAM-dependentO-methyltransferasefrompathogenicbacteriumLeptospirainterrogans.
J.
Struc.
Biol.
2007;159(3):523-8.
5.
XLi,ZWei,MZhang,XPeng,GYu,MTeng,WGong.
CrystalstructuresofE.
colilaccaseCueOatdifferentcopperconcentrations.
Biochem.
Bioph.
Res.
Co.
.
2007;354(1):21-66.
ZCheng,LSun,JHe,WGong.
Crystalstructureofhumanmu-crystallincomplexedwithNADPH.
ProteinSciences,2007;16(2):329-357.
CZhang,LLiu,HXu,ZWei,YWang,YLin,WGong.
CrystalstructuresofhumanIPPisomerasenewinsightsintothecatalyticmechanism.
JMolBiol.
2007;366(5):1437-46153、江涛组2007年我们一共完成了包括神经生长因子,神经营养因子与其受体P75NTR复合物,P2蛋白,Mastoparan以及钙调蛋白-靶肽复合物等五种蛋白的晶体结构解析工作,并获得多种重要蛋白的晶体.
1、神经营养因子对于神经系统的发育具有重要的意义.
研究发现,其细胞表面受体之一p75NTR与神经营养因子的直接相互作用与神经系统退行性疾病关系密切.
但是二者间的相互作用方式一直存在争议.
我们最近完成了神经营养因子NT-3与75NTR2:2复合物2.
8分辨率晶体结构解析,揭示了神经营养因子与其受体P75NTR天然的结合模式,因此具有重要的科学意义.
2、Mastopran是蜂毒中一种具有多种生物学功能的多肽,我们利用晶体学手段解析了这种多肽1.
2分辨率的晶体结构.
通过与核磁结构比较,并结合生物化学实验的结果,我们对其结构功能关系进行了论述.
3、P2蛋白是外周髓鞘中的重要蛋白.
迄今其功能未知,但有实验证明P2能诱导实验性过敏神经炎(EAN),可作为研究人类Guillain-Barré综合症的模型.
我们解析了猪的硬脊髓根部神经中的P2,获得分辨率为1.
8的晶体结构.
根据电子密度图在P2结合口袋处发现其天然脂类配体,初步推断为γ-亚麻酸(GLA,18:3n-6).
这一发现为P2的功能研究提供了新的思路.
4、膜受体Fas与其配基FasL的相互作用与人类癌症的发生关系密切以往的研究表明,钙调素与Fas死亡结构域之间存在着直接的相互作用.
我们最近完成了CaM与Fas肽复合物的晶体结构解析,该结构有助于人们深入了解CaM与Fas受体间的相互作用方式.
3、Prof.
JiangT.
In2007,wedeterminedfiveproteinstructuresincludingnervegrowthfactor,NT3-P75complex,Mastoparan,CalmodulincomplexandP2protein.
1)NTsarekeyfactorsforthedevelopmentandmaintenanceofthemammaliannervoussystem.
ThebindingofNTstothep75NTRreceptorinduceseithercellsurvivalorapoptosis.
p75NTRalsoplaysaveryimportantroleinAlzheimer'sdisease,andtumordevelopmentandmetastasis.
Inspiteofthisknowledge,thefundamentalmechanismofinteractionbetweenp75andNTsisnotclear.
Herewereportthecrystalstructureofa2:2symmetricalcomplexofNT-3andp75NTRat2.
8Aresolution.
Thisstructurerevealedthedetailedinteractionmodebetweenp75withNT-3.
Thisstructureprovidesabasistounderstanddiversep75NTRfunctionsandfacilitatethetherapeuticutilityofneurotrophinsasclinicalagents.
2)Mastoparansarethemajorpeptidesinsocialwaspvenomsandpossessavarietyofbiologicalactivities.
HerewereportthefirstcrystalstructureofmastoparanfromPolistesjadwagae(MP-PJ)at1.
2resolution.
Togetherwithbiochemicalresults,weproposethattheinteractionsbetweenmastoparanmoleculesserveanimportantroleinformingtheα-helixconformationwhichishighlyrelatedtotheirbiologicalactivities.
3)TheP2proteinisasmallbasicproteinfoundinperipheralmyelin.
Itsfunctionhasnotbeenestablished,butitisknownthatP2couldinduceexperimentalallergicneuritis(EAN)inrats,amodelofGuillain-Barrésyndromeinhumans.
StructureofP2proteinpurifiedfromintraduralspinalrootsofpigwasdeterminedto1.
8.
Thestructurecontainssomeextraelectrondensityinthebindingcavity.
ItisverylikelythatthisisthedensityofP2protein'snaturallipidligand.
Thedensityisquitesuitableforthemolecularofγ-linolenicacid(GLA,18:3n-6),whichwasnotreportedpreviously.
发表文章:LiuSQ,WangF,TangL,GuiWJ,CaoP,LiuXQ,PoonWS,ShawPC.
CrystalstructureofmastoparanfromPolistesjadwagaeat1.
2angstromresolution.
JOURNALOFSTRUCTURALBIOLOGY2007;160(1):28-34164、梁栋材组1)在甘油的代谢过程中,甘油磷酸二酯磷酸二酯酶(GDPD)催化甘油磷酸二酯,水解成一个醇类和一个三磷酸甘油.
它的产物参与到很多的生化途径中,起到重要的作用.
我们解析了来自于T.
tengcongensis的GDPD(ttGDPD)的1.
9埃的结构.
ttGDPD单体分子通过一个分子间的二硫键以及其他一些氢键结合形成二体,二体很可能是作为它的功能单位.
通过对ttGDPD同源蛋白的一级序列比对以及三维结构叠合,我们在活性位点周围确定了一些保守残基,可能和ttGDPD的催化机制有密切关系.
通过对这些残基的单点突变研究,i)我们首次确定了GDPD是一个金属依赖酶;ii)确定了ttGDPD的活性残基,并提出了ttGDPD的可能的催化机制;iii)根据同源蛋白1zcc的结构中活性位点结合的硫酸根离子,通过把它模拟成GDPD底物中的磷酸半族,分析了ttGDPD底物分子甘油磷酸二酯分子和ttGDPD的实际结合情况.
2)细菌通过趋化系统来控制自身运动方向,从而对有益或有害化学信号的梯度变化做出响应.
两组跨膜蛋白复合物参与了细菌对外界环境和自身生理信号的感应、增益和反馈控制.
CheW是第一组复合物中的重要因子.
它与CheA和MCP结合,在细胞极点上对稳定MCP-CheA的结合起到了至关重要的作用.
在2.
2分辨率下,我们运用分子置换法解析了来源于腾冲嗜热菌中蛋白TtCheW的三维结构,结构分析发现在CheW上存在一个保守motif"NxxGxIxP",对CheW-CheA的结合起了重要作用.
我们根据已有的CheW单点突变实验和对TtCheW分子中蛋白质相互作用区域预测,推测TtCheW与MCP的结合区域是位于复合物结构模型中TtCheW和CheAP4domain结合所产生的凹槽内.
3)完成了二种重要功能酶的三维结构研究.
①利用SAD方法解析了Tfdx蛋白的2.
15的三维结构.
Tfdx属于具有类β-内酰氨酶结构域的金属水解酶家族,该蛋白折叠形式为αβ/βα形式,β-内酰氨酶折叠结构域的金属结合位点在水解共价键的过程中起作用.
文章已投送国际期刊.
②利用SIRAS方法解析了来自生防芽孢杆菌的甘露聚糖酶BCman1.
45的三维结构.
通过功能研究与结构分析揭示了BCman催化特性和耐热性.
4、Prof.
LiangD.
C.
1)Glycerophosphodiesterphosphodiesterasecatalyzesthehydrolysisofaglycerophosphodiestertoanalcoholandglycerol3-phosphateinglycerolmetabolism.
Ithasanimportantroleinthesynthesisofavarietyofproductsthatparticipateinmanybiochemicalpathways.
WereportthecrystalstructureoftheThermoanaerobactertengcongensisGDPD(ttGDPD)at1.
9resolution.
ThettGDPDdimerwithanintermoleculardisulfidebridgeandtwohydrogenbondsisconsideredasthepotentialfunctionalunit.
Toelucidatethecatalyticmechanism,weusedsite-directedmutagenesistomakeaseriesofconvertedresidues.
WehavecharacterizedttGDPDasametalion-dependentenzyme,forthefirsttime.
Wehavealsoidentifiedtheresiduesparticipatinginthecatalysis,andweproposethecatalyticmechanismofttGDPDtoo.
ThereisasulfateionintheactivesiteofhomologousstructureGDPDfromA.
tumefaciens(1zcc),whichisequivalenttothephosphatemoietyofthesubstrate,allowingustostudythetruesubstrate-bindingmodeofttGDPD.
2)Chemotaxisenablesbacteriatocontroltheirmovementsinresponsetogradientsofbenecialandtoxicchemicals.
Twotransmembraneproteincomplexesfrombacteriaexhibitremarkablesensitivity,throughgainandfeedbackcontrol,totheexternalenvironmentandinternalphysiology.
CheWisanessentialcomponentoftherstcomplex.
ItinteractswithbothCheAandthecytoplasmicdomainofMCPandlinkstheiractivities.
ThecrystalstructureofthescaffoldingproteinCheWfromThermoanaerobactertengcongensis(TtCheW)issolvedwitharesolutionat2.
2Ausingmolecular17replacement.
Basedonthecrystalstructure,wefoundthattheconservedmotif''NxxGxIxP''fromCheWplaysanimportantroleinCheAbinding.
ThecoincidenceofthereportedmutationsitesrelatedtoCheW–MCPbinding,andthepredictedproteininteractionregionwithintheTtCheWmolecule,suggestthatCheW–MCPbindingsiteslieinthegroove-shapedareabetweenTtCheWandtheCheAP4domainwithintheassembledmodel.
3)Solvedthecrystalstructureoftwoimportantfunctionenzymes.
①Wehavesolvedthe2.
15crystalstructureofTfdxusingsingleSAD.
Tfdxbelongtoametalhydrolyzefamily,whichhavetheβ-inneracylammoniasedomain.
TheTfdxmoleculeexhibitsfoldofαβ/βα.
Itsmetalbindingsiteplaysanimportantroleinthecourseofhydrolyzescovalentbond.
Articletobecontributed.
②Wehavesolvedthe1.
45crystalstructureofBCmanusingSIRAS.
StructureandfunctionstudiesrevealedCatalyticSubstrateSpecificityandThermostabilityDisplayed.
Articletobecontributed.
发表文章:WangYao,LiangShi,Dong-CaiLiang.
CrystalstructureofscaffoldingproteinCheWfromthermoanaerobactertengcongensis.
BiochemicalandBiophysicalResearchCommunications2007;361:1027-1032.
185、刘迎芳组本课题组今年的工作进展主要集中在凋亡细胞的吞噬和泛素蛋白酶体两大途径.
在线虫中凋亡细胞的吞噬主要由两条信号通路介导,第一条为CED-14/CED-1/CED-6/CED-7,第二条信号通路为CED-2/CED-5/CED-10/CED-12.
目前我们已经解析了CED-14和CED-2的晶体结构,它们都在凋亡细胞的吞噬方面起着非常重要的作用,结构的解析,为疾病的研究提供了新的思路.
底物泛素化主要包括E1泛素激活酶,E2泛素转移酶,E3泛素连接酶和近年发现的多泛素连接因子E4.
DCN1是一种参与泛素-蛋白酶体途径泛素化修饰过程的蛋白,结构表明它可能是E3连接酶酶复合物的一种结合接头蛋白,用以连接泛素化蛋白与底物蛋白.
DOA1是E4家族成员中的一员,它能与泛素和CDC48直接结合,从而帮助CDC48招募泛素来行使它的功能,从结构上分析,它在蛋白质泛素降解途径中起到非常重要的作用,阐明了泛素蛋白质降解途径中泛素来源问题.
另外,本课题组承担了国家自然科学基金重大项目"禽流感病毒主要蛋白质的结构和功能研究",并获得了其中RNA聚合酶亚基的晶体,结构正在进一步解析中.
5、Prof.
LiuY.
F.
Ourlabmainlyfocusedonthecelldeathengulfmentandubiquitin-proeasomepathwaythisyear.
1,Dyingcellsengulfmentandagingresearch:TwosignaltransductionpathwaysthatactredundantlytocontrolengulfmentinCeleganshavebeenidentifiedinthepastreports.
Componentsofthecell-corpserecognitionsystemofonepathwayincludestheCED-14/CED-1/CED-6/CED-7;ThesecondpathwaycontainsCED-2/CED-5/CED-12.
NowwehavesolvedthestructuresofCED-14andCED-2.
Thestructuresoftheseproteinsimplicatetheirmechanismsforcellcorpseengulfmentprocesses.
Proteinubiquitination,aprocessesthatcovalentlylinkubiquitinorubiquitin-likeproteinstoproteinsubstrates,involveinE1(ubiquitinactivatingenzyme),E2(ubiquitinconjugatingenzyme),E3(ubiquitin-proeinligase)andE4(ubiquitinelongatingfactor).
DCN1participatesinmodificationofproteinsubstratesbyNedd8inubiquitin-proeasomepathway.
ThestructureofDCN1illustratesthatDCN1couldbeascaffoldproteininanE3ligasecompelx.
DOA1isamemberofE4,whichcanbindubiquitinandCDC48tohelpCDC48recruitubiquitintofacilitatetheirfunction.
DOA1playsakeyroleduringubiquitindependentproteindegradationbasedonstructureanalysesandisthemajorsourceofubiquitinduringubiquitindependentproteindegradationprocess.
WealsotookpartinthestructuralandfunctionalstudyofthemainproteinofavianinfluenzavirusandobtainedthecrystalofRNApolymerasesubunitandstructuredeterminationisundergoing.
发表文章:YangX,ZhouJ,SunL,WeiZ,GaoJ,GongW,XuRM,RaoZ,LiuY.
StructuralbasisforthefunctionofDCN-1inproteinNeddylation.
JBiolChem.
2007;282(34):24490-4.
196、刘志杰组本课题组过去的一年中,在所各级领导的关怀和结构生物学中心、及平台各位老师的帮助下,带领课题组成员们顺利实施了获得资助的几个研究课题并且进展顺利,取得了一些阶段性的研究成果,2007年共发表研究论文12篇,包括NatStrucMolBiol,Cell,Proteins等.
同时又争取到了5项研究基金,总之各项工作稳步进行.
在科研经费申请方面,获得我院"知识创新工程重要方向项目"和与韩国KIST合作的资助项目各一项;与英国Leeds大学的StephenBaldvin教授合作获得访问学者交流基金一项;从院国际合作局获得与俄罗斯科学院生物物理所EugeneVysotski教授合作的国际交流资助一项;我组副研究员NeilShaw获得国家自然科学青年基金资助一项.
在合作与交流方面,积极参加了多个结构生物学方面的国际会议并作邀请报告.
接待多位海外科学家的到访.
主办了"InternationalSymposiumonSynchrotronRadiationandBiology".
在研究所建设方面,积极参与平台二期建设,负责蛋白质平台二期"自动化克隆和小规模表达可溶性筛选机器人"和"蛋白质晶体生长机器人"筹备工作,该工作已基本完成.
在新的一年中,本人将继续带领研究组成员在院和研究所的政策方针指导下,进一步加强自主创新能力和核心竞争力的培养,争取取得更大成果.
6、Prof.
LiuZ.
J.
In2007,ourteammademanysignificantachievements.
1.
Papers:12researchpaperswerepublishedinhighimpactfactorjournals,includingNat.
Struc.
Mol.
Biol.
2007,14(8),779-84;Cell2007,129,747-759;Proteins2007(1),263-2672.
Grants:①fundfromCAS,②fundofKISTproject,③fundfromCAS-RAS,④fundfromNSFC.
3.
Awards:DoctorNeilShawWontheTopPrizeatthe6thTICPSinternationalconference.
4.
Internationalcooperation:①organizedInternationalSymposiumonSynchrotronRadiationandBiology,②attendedseveralinternationalconferencesonStructuralBiology.
5.
Researchplatformdevelopment:Automatedcloning&small-scaleexpressiontestingusingrobotics,proteincrystallizationatnanoliterscaleusingroboticsimplemented.
发表文章:1.
Shaw,N.
,Zhao,M.
,Cheng,C.
,Xu,H.
,Saarikettu,J.
,Li,Y.
,Da,Y.
,Yao,Z.
,Silvennoinen,O.
,Yang,J.
,Liu,Z.
J*.
,Wang,B.
C.
&Rao,Z.
Themultifunctionalhumanp100protein'hooks'methylatedligands.
NatStructMolBiol,2007;14(8):779-84.
(Correspondence)2.
Shaw,N.
,Tempel,W.
,Chang,J.
,Yang,H.
,Cheng,C.
,Ng,J.
,Rose,J.
,Rao,Z.
,Wang,B.
C.
&Liu,Z.
J*.
(2007b)Crystalstructuresolutionofaparb-likenucleaseatatomicresolution.
Proteins,Doi:10.
1002/prot.
21641.
(Correspondence)3.
Shaw,N.
,Cheng,C.
,Tempel,W.
,Chang,J.
,Ng,J.
,Wang,X.
Y.
,Perrett,S.
,Rose,J.
,Rao,Z.
,Wang,B.
C.
&Liu,Z.
J*,.
(NZ)CH.
.
.
OContactsAssistCrystallizationofaParB–likeNuclease,BMCStrucBiol.
2007;7:46.
(Correspondence)4.
Shimada,A.
,Niwa,H.
,Tsujita,K.
,Suetsugu,S.
Kyoko,H.
S.
,Akasaka,R.
,Nishino,Y.
,Toyama,M.
,Chen,L.
,Liu,Z.
J,Wang,B.
C.
,Yamamoto,M.
,Terada,T.
,Miyazawa,A.
,Shirouzu,M.
,Tanaka,A.
,Sugano,S.
,Takenawa,T.
,andYokoyama,S.
CurvedEFC/F-BARdomaindimersarejoinedendtoendintoafilamentformembraneinvaginationinendocytosis.
Cell.
2007;129:747-759.
5.
Liu,Z.
J*.
,Chen,H.
,Shaw,N.
,Hopper,S.
L.
,Chen,L.
,Chen,S.
,Cerniglia,C.
E.
,andWang,B.
C.
,CrystalstructureofanaerobicFMN-dependentazoreductase(AzoA)fromEnterococcusfaecalis.
Arch20BiochemBiophys.
2007;463,68-77.
(Correspondence)6.
Das,A.
,Fu,Z.
Q.
,Tempel,W.
,Liu,Z.
J.
*,Chang,J.
,Chen,L.
,Lee,D.
,Zhou,W.
,Xu,H.
,Shaw,N.
,Rose,J.
P.
,Ljungdahl,L.
G.
,andWang,B.
C.
,CharacterizationofacorrinoidproteininvolvedintheC1metabolismofstrictanaerobicbacteriumMoorellathermoacetica.
Proteins.
2007;67:167-176.
(Correspondence)7.
TakuyaB.
Hiyama,MinZhao,YuKitago,MinYao,Shun-ichiSekine,TakahoTerada,ChizuKuroishi,Liu,Z.
J.
,JohnP.
Rose,SeikiKuramitsu,MikakoShirouzu,NobuhisaWatanabe,ShigeyukiYokoyama,IsaoTanaka,andWang,B.
C.
.
StructuralbasisofCoArecognitionbythePyrococcussingle-domainCoA-bindingproteins.
JStructFunctGenomics,2007;7:119-129.
8.
Gerwe,B.
,Kelley,L.
L.
,Dillard,B.
D.
,Lai,T.
,Liu,Z.
J.
,Tempel,W.
,Chen,L.
,Habel,J.
,Lee,D.
,Jenney,F.
E.
,Jr.
,Sugar,F.
J.
,Richardson,J.
S.
,Richardson,D.
C.
,Newton,M.
G.
,Wang,B.
C.
,Adams,M.
W.
&Rose,J.
P.
Structuralandtranscriptionalanalysesofapurinenucleotide-bindingproteinfrompyrococcusfuriosus:Acomponentofanovel,membrane-boundmultiproteincomplexuniquetothishyperthermophilicarchaeon.
JStructFunctGenomics,2007;8(1),1-10.
9.
Kondo,N.
,Nakagawa,N.
,Ebihara,A.
,Chen,L.
,Liu,Z.
J.
,Wang,B.
C.
,Yokoyama,S.
,Kuramitsu,S.
Masui,R.
StructureofdNTP-inducibledNTPtriphosphohydrolase:insightintobroadspecificityfordNTPsandtriphosphohydrolase-typehydrolysis.
ActaCrystD2007;63,230-9.
10.
Kanaujia,S.
P.
,Ranjani,C.
V.
,Jeyakanthan,J.
,Baba,S.
,Chen,L.
,Liu,Z.
J.
,Wang,B.
C.
,Nishida,M.
,Ebihara,A.
,Shinkai,A.
,Kuramitsu,S.
,Shiro,Y.
,Sekar,K.
,andYokoyama,S.
Crystallizationandpreliminarycrystallographicanalysisofmolybdenum-cofactorbiosynthesisproteinCfromThermusthermophilus.
ActaCrystF2007;63,27-29.
11.
Kelley,L.
L.
;Dillard,B.
D.
;Tempel,W.
;Chen,L.
;Shaw,N.
;Lee,D.
;Newton,M.
G.
;Sugar,F.
J.
;Jenney,F.
E.
,Jr.
;Lee,H.
S.
;Shah,C.
;Poole,F.
L.
,3rd;Adams,M.
W.
;Richardson,J.
S.
;Richardson,D.
C.
;Liu,Z.
J.
;Wang,B.
C.
;Rose,J.
StructureofthehypotheticalproteinPF0899fromPyrococcusfuriosusat1.
85resolution.
ActaCrystF2007;63,549-552.
12.
Cacciapuoti,G.
,Porcelli,M.
,Moretti,M.
A.
,Sorrentino,F.
,Concilio,L.
,Zappia,V.
,Liu,Z.
J.
,Tempel,W.
,Schubot,F.
,Rose,J.
P.
,Wang,B.
C.
,Brereton,P.
S.
,Jenney,F.
E.
&Adams,M.
W.
Thefirstagmatine/cadaverineaminopropyltransferase:Biochemicalandstructuralcharacterizationofanenzymeinvolvedinpolyaminebiosynthesisinthehyperthermophilicarchaeonpyrococcusfuriosus.
JBacteriol,2007;189(16),6057-67217、王大成组1.
2006-2007年完成4种蛋白质及复合物的晶体结构研究,2007年内已发表和被接受发表论文2篇.
2.
2007年内完成动物病原菌三型分泌系统蛋白质效应物SpvC及其活性多肽底物三维结构及其与功能关系的研究,发现全新的蛋白质结构,阐明SpvC与致病通路中关键激酶MAPK相互作用,进而阻断宿主天然免疫应答的机理.
由于这类病原菌效应物同时存在于动植物中,因而具有重要意义.
有关论文已发表在MolecularCell.
3.
完成3种Cul3介导的泛素连接酶复合物的体外构建,其中2种已获得可溶表达和纯化样品,一种获得初步结晶.
7、Prof.
WangD.
C.
In2006-2007,fourcrystalstructureshavebeendeterminedand2paperspublishedoracceptedin2007(seepaperlist).
Amongothers,crystalstructuresofSpvC,aneffectorofTTSSofbacterialpathogens,anditscomplexwithphosphopeptidesubstrate,revealedanovelproteinstructuraltypeandprovidesinsightsintothemolecularmechanismoftheinactivationofthepathogenicMAPK.
TheresultshavebeenpublishedinMolecularCell.
Besides,3humanCul3ubiquitinligasecomplexeshasbeenconstructed.
Amongthem2recombinantcomplexeshavebeenexpressedandpurified,onecrystallizedpreliminary.
发表文章:YongqunZhu,HongtaoLi,ChengzuLong,LiyanHu,HaoXu,LipingLiu,SheChen,Da-ChengWang,FengShao.
StructuralInsightsintotheEnzymaticMechanismofthePathogenicMAPKPhosphothreonineLyase.
MolecularCell,2007;28:899-91322(二)蛋白质功能与折叠原理研究1、陈畅组细胞的氧化还原调控失衡与衰老、神经退行性疾病、炎症、糖尿病等都密切相关,其分子机制有待阐明.
我们的研究方向为细胞氧化还原调控的分子机制:氧化还原依赖的蛋白质翻译后修饰在细胞命运和疾病发生中的作用.
重点研究一氧化氮对蛋白巯基的亚硝基化修饰及作用.

1.
致力于蛋白质巯基亚硝基化方法学的原始创新:发现了维生素C在亚硝基化检测过程中引起假阳性信号;解决了高通量检测蛋白亚硝基化方法中表面活性剂对质谱的干扰和实验重复性差的问题;创建了蛋白亚硝基化的胶内快速直接可见检测方法;已建立高通量定量组学检测内源亚硝基化方法.
2.
发现一氧化氮引起的蛋白质亚硝基化修饰失衡在细胞死亡中的关键作用,并首次阐述了蛋白质亚硝基化修饰与细胞氧化还原状态的定量依赖关系.
3.
发现一氧化氮通过蛋白亚硝基化修饰诱导核酸内切酶APE1-REF1核输出,首次揭示了一氧化氮通过蛋白亚硝基化修饰特异机制实现对细胞蛋白核输入和核输出系统的调控.
一氧化氮通过蛋白质巯基亚硝基化修饰调控SUMO修饰E3连接酶PIAS3稳定性,首次揭示了蛋白亚硝基化修饰、泛素化修饰及SUMO化修饰三种翻译后修饰的相互作用.
4.
在研究单个蛋白亚硝基化对其功能调控的基础上,结合本课题组建立的蛋白亚硝基化修饰的定量组学方法和系统生物学思想,进行多靶点蛋白亚硝基化修饰作用网络的分析和动态变化研究.
1、Prof.
ChenC.
Weareinterestedininvestigatingthemechanismandthecellulareffectsoftheredox-basedpost-translationalmodificationofproteins,particularlyontheS-nitrosylationofproteinsinducedbynitricoxide.
Someprogressin2007isasfollowing:Wefoundthatnitricoxidecausesglobalhyposumoylationinmammaliancells,promotedPias3degradationbyfacilitatingitsinteractionwithtripartitemotif-containing32(Trim32),aubiquitinE3ligase.
ThisstudyrevealsanovelcrosstalkbetweenS-nitrosation,ubiquitination,andsumoylation,whichmaybecrucialforNO-relatedphysiologicalandpathologicalprocesses.
WefoundthatnitricoxidecontrolsnuclearexportofAPE1/Ref-1throughS-nitrosationofCysteines93and310.
OurfindingprovidedthefirstevidencethatnitrosativestressregulatedAPE1bysubcellulartranslocationthroughS-nitrosationmechanism.
Weproposedanovelmechanismunderlyingthesusceptibilityofneuronalcellstonitricoxide:theoccurrenceandregulationofproteinS-nitrosylationisthecheckpoint.
Wedevelopedadetergent-freebiotinswitchmethodandcombinedwithLC-MS/MStoidentifytheS-nitrosylatedtargets.
TheLC-MSperformancefortheproteomicanalysisofS-nitrosylatedproteinswasgreatlyamelioratedandtherepeatabilityofexperimentswasgreatlyimproved.
Furthermore,theamountofsamplewasalsosignificantlyreduced.
发表文章:1.
JingQu,Guang-HuiLiu,KaiyuanWu,PeiweiHan,PengWang,JiangmeiLi,XuZhang,ChangChen*,NitricOxideDestabilizesPias3andRegulatesSumoylation.
PLoSONE,2007;2(10):e1085.
232.
JingQu,Guang-HuiLiu,BoHuang,ChangChen*NitricoxidecontrolsnuclearexportofAPE1/Ref-1throughS-nitrosationofCysteines93and310.
NucleicAcidsResearch,2007;35:2522-25323.
JieHe,TiepengWang,PengWang,PeiweiHan,ChangChen*,Anovelmechanismunderlyingthesusceptibilityofneuronalcellstonitricoxide:theoccurrenceandregulationofproteinS-nitrosylationisthecheckpoint.
JournalofNeurochemistry,2007;102:1863–18744.
ShaojinDuan,ChangChen*,S-nitrosylation/DenitrosylationandApoptosisofImmuneCells.
Cellular&MolecularImmunology.
2007;Vol.
4(5):353-358.
242、柯莎组在2007年度完成的工作主要有五方面:1)Ure2prion化的分子机制:N端和C端结构域分别在prion化中的作用酵母prion蛋白Ure2在体内和体外都能形成淀粉样结构,其N端的prion结构域(PrD)和C端部分区域被发现在体内能影响Ure2的prion化.
为了揭示Ure2prion化的机制,我们已经研究了一系列PrD不同程度缺失的Ure2突变体的性质,发现PrD在纤维化的成核过程中有重要作用,C端区域决定Ure2的折叠性质.
我们也进行Ure2折叠的动力学和热力学分析,发现Ure2至少存在三个折叠中间态,目前我们正在研究一系列C端缺失突变体的折叠机制.
2)Ure2的酶学分析我们首次证明了天然态和纤维化Ure2都具有谷光甘肽依赖性的过氧化物酶活性.
为了进一步发现其反应机制和确认催化活性的必需残基,我们正在进行一系列相关突变体的构建以及功能与结构关系的研究.
3)淀粉样聚集物的细胞毒性研究培养不同种类的哺乳动物细胞,加入天然态Ure2及不同种类的Ure2聚集体,研究Ure2及其成纤维不同阶段聚集体的细胞毒性.
原初纤维的细胞毒性最大,其次是成熟纤维,天然态的Ure2几乎没有细胞毒性.
分子伴侣对Ure2聚集体细胞毒性的影响正在研究中.
4)不同酵母来源Ure2的对比研究有研究表明Saccharomyces.
paradoxusUre2p(SpUre2p)SacharomycescerevisiaeUre2p(ScUre2p)有很高的同源性,但体内实验表明,SpUre2p并没有prion形成的能力.
为此,我们和法国Cullin教授的实验室合作,分别从体外和体内做实验来寻找原因.
我们的体外实验证明,SpUre2p同ScUre2p一样,在体外都有着很强的成纤维的能力,但SpUre2p的纤维有着更好的机械强度.
于是我们推测,可能正是因其高机械强度,SpUre2p的纤维不容易被打断产生种子传播prion,导致prion的丧失.
5)分子伴侣与Ure2的相互作用在细胞内影响prion化的因素包括分子伴侣Hsp70、Hsp40和Hsp104.
这些分子伴侣已被纯化并用于探索他们对Ure2折叠和纤维化的影响.
最近的结果表明Hsp40-Ydj1能和Ure2直接相互作用,并且特异性的在Ure2成纤维的早期抑制Ure2的纤维形成.
2、Prof.
Perrett,S.
ProteinMisfoldingandDisease1)MolecularmechanismofUre2prionformationTheyeastprionproteinUre2formsamyloid-likestructureinvivoandinvitro.
MutantsineithertheN-terminalpriondomain(PrD)ortheC-terminalregionofUre2arefoundtoaffectprionformationinvivo.
Inordertoshedlightonthemechanismofprionformation,wehavestudiedthepropertiesofaseriesofPrDdeletionmutants.
WefoundthattheN-terminalPrDplaysanimportantroleinthenucleationofamyloid-likestructure,whilethefoldingpropertiesofUre2aredeterminedbytheC-terminalregion.
WehavealsocarriedoutadetailedkineticandthermodynamicanalysisofUre2folding,identifyingatleastthreefoldingintermediates,andwearecurrentlystudyingthefoldingmechanismofaseriesofC-terminaldeletionmutants.
2)EnzymaticactivityofUre2WehavedemonstratedthatUre2possessesglutathione-dependentperoxidaseactivity.
Interestingly,wefoundthatperoxidaseactivityismaintainedinamyloid-likefibrilsofUre2.
Inordertogainfurther25insightintothereactionmechanismandtoidentifytheresiduesrequiredforcatalyticactivity,wearecurrentlycarryingoutmutagenesisstudies.
Basedoninspectionofthepublishedcrystalstructure,andsequencealignmentsofUre2withrelatedenzymes,wehavemutatedresiduesthatappeartobeimportantforsubstratebindingoractivation.
Themutantshavebeenpurifiedandtheirenzymekineticscharacterized.
ApictureisnowemergingoftheresiduesimportantfortheenzymeactivityofUre2.
3)IdentifyingthetoxicspeciesintheprocessofamyloidformationWehaveisolatedintermediatesthatappearatdifferentstagesduringtheprocessofUre2amyloidfibrilformationandtestedtheiraffectonmammaliancellsinculture.
Wefoundthatbothsmallaggregatesandmaturefibrilswereabletoentermammaliancells,butonlyearlyaggregationintermediatesweretoxictothecells.
4)ConservationofUre2PrionBehaviourinDifferentYeastSpeciesWhileUre2intheyeastspeciesSaccharomycescerevisiae(ScUre2)behavesasaprion,itshomologue(SpUre2)inthecloselyrelatedyeastS.
paradoxus,doesnot.
Inordertoinvestigatethestructuralbasisforthis,wecollaboratedwithProf.
C.
Cullin,Bordeaux,France,tocomparethepropertiesoftheproteinsinvitroandinvivo.
Surprisingly,wefoundthatSpUre2,likeScUre2,readilyformsamyloid-likefibrilsinvitro.
Further,thebiochemicalpropertiesofthetwoproteinsareidentical.
However,weobservedgreaterresistancetofragmentationforSpUre2fibrils.
TheresultssuggestthatthefailureofSpUre2toactasaprionstemsfromitsinabilitytopropagatetransmissibleprionsinvivo,ratherthanafundamentaldifferenceinamyloidstructure.
5)InteractionofchaperoneswithUre2FactorsthataffectUre2prionformationinthecellincludethemolecularchaperonesHsp70,Hsp40andHsp104.
ThesechaperoneshavebeenpurifiedandweareinvestigatingtheireffectuponfoldingandfibrilformationofUre2.
AdetailedcharacterizationoftheinteractionwithHsp40indicatesthatitinhibitsfibrilformationbybindingtothenativestateofUre2priortotheonsetofoligomerisation.
发表文章:1.
Gao,L.
,Zhuang,J.
,Nie,L.
,Zhang,J.
,Gu,N.
,Wang,T.
,Perrett,S.
&Yan,X.
Intrinsicperoxidase-likeactivityofferromagneticnanoparticles.
NatureNanotechnology2007;2,577-583.
2.
Shaw,N.
,Tempel,W.
,Chang,J.
,Ng,J.
,Wang,X.
Y.
,Perrett,S.
,Rose,J.
,Rao,Z.
,Wang,B.
C.
,Liu,Z.
J.
(NZ)CH.
.
.
OContactsAssistCrystallizationofaParB-likeNuclease.
BMCStructuralBiology2007;7,46.
3.
Shi,Y.
,Fan,D.
J.
,Li,S.
X.
,Zhang,H.
J.
,Perrett,S.
andZhou,J.
M.
IdentificationofapotentialhydrophobicpeptidebindingsiteintheC-terminalarmoftriggerfactor.
ProteinScience2007;16,1165-1175.
4.
Lian,H.
Y.
,Zhang,H.
,Zhang,Z.
R.
,Loovers,H.
M.
,Jones,G.
W.
,Rowling,P.
,Itzhaki,L.
S.
,Zhou,J.
M.
&Perrett,S.
Hsp40interactsdirectlywiththenativestateoftheyeastprionproteinUre2andinhibitsformationofamyloid-likefibrils.
J.
Biol.
Chem.
2007;282,11931-11940.
5.
Nie,C.
L.
,Wang,X.
S.
,Liu,Y.
,Perrett,S.
&He,R.
Q.
.
Amyloid-likeaggregatesofneuronaltauinducedbyformaldehydepromoteapoptosisofneuronalcells.
BMCNeuroscience.
2007;8,9.
6.
Immel,F.
,Jiang,Y.
,Wang,Y.
Q,Marchal,C.
,MailletL.
,Perrett,S.
&Cullin,C.
InvitroanalysisofSpUre2p,aprionrelatedprotein,exemplifiestherelationshipbetweenamyloidandprion.
J.
Biol.
Chem.
2007;282,7912-7920.
263、秦燕组克隆表达并纯化出大肠杆菌LepA和EF-G的全长与缺失体蛋白,测定了它们的GTPase活性,正在检测它们的分子伴侣活性;已获得LepA全长蛋白的晶体,正在优化条件;发酵得到大量对数期大肠杆菌菌体,并通过区带密度梯度离心方法制备出足够多的重组核糖体,测定其具有体外活性;制备并纯化得到LepA蛋白与核糖体L2蛋白的多克隆抗体,正在制备LepA蛋白的单克隆抗体;合成特定的氨酰tRNA与mRNA,构建体外翻译体系,为进一步实验提供了良好基础;通过大肠杆菌野生型与LepA缺陷型在极端条件下的生长曲线对LepA蛋白的功能作出提示,正在对LepA蛋白进行细胞定位;正在制备敲除lepA同源基因的果蝇;表达纯化得到大肠杆菌RRF蛋白,蔗糖密度梯度离心获得大肠杆菌多聚核糖体,准备进行多聚核糖体breakdown实验.
3、Prof.
QinY.
Escherichiacolifull-lengthandtruncatedprotensofLepAandEF-Ghavebeencloned,expressedandpurified,andtheirGTPaseactivityhasalsobeentested.
Thechaperoneactivityofthistwoproteinsaswellastheirtruncationsareundertest.
WehaveobtainedthecrystalofLepA,butitsgrowthconditionisstillinoptimization.
PlentyofrecombinantribosomehasbeenextractedfromE.
coliinexponentialphaseandhasbeenseparatedandpurifiedbyzonaldensitygradientcentrifugation.
Ribosomeactivityinvitrohasbeenascertained.
PolyclonalantibodiesofLepAandribosomeproteinL2areavailable.
LepAmonoclonalantibodyisunderpreparation.
Specificaminoacyl-tRNAandmRNAhavebeenpreparedandinvitrotranslationsystemhasbeenconstructed,whichprovidesaconvenientfoundationforfuturestudy.
ThegrowthcurveresultsofE.
coliwidetypeandlepAknock-outstrainshavegivenussomecluesofLepAfunctioninvivounderextremegrowthconditions.
LocalizeLepAinvivo.
AndwearetryingtoknockoutthehomologousgeneoflepAinfruitfly.
ProteinRRFofE.
colihasbeenexpressedandpurifiedandpolysomehasalsobeenseparated,sothatthebreakdownassayofpolysomeisgoingtobecarriedout.
274、王志珍组1.
α-synuclein(AS)在大肠杆菌的转运α-synuclein(AS)是帕金森氏病患者大脑神经元中淀粉样斑块的主要成分.
确认了在大肠杆菌表达的无信号肽的AS转运到周质腔定位.
鉴定其C端99-140是与转运相关的关键序列,可能起到类似信号肽的作用;N端1-60序列对其转运没有贡献.
发表在2007年J.
Bacteriol.
.
2.
ERp44的晶体结构解析人内质网滞留蛋白ERp44是PDI家族成员,也是ER内蛋白质质量控制的重要成员.
解析了2.
6埃分辨率人ERp44的晶体结构,登录PDB(ID:2r2j).
这是迄今唯一获得的人PDI家族成员全长分子的晶体结构.
发现柔性的C末端尾巴挡住结合底物蛋白的疏水口袋和CRFS基序周围的疏水表面.
切除C端使酶活性、分子伴侣活力和结合内源性蛋白能力明显增强.
4、Prof.
WangC.
C.
1.
Translocationofα-SynucleinexpressedinEscherichiacoli.
α-synuclein(AS)isamajorcomponentofLewybodiesinParkinson'sdisease.
WereportforthefirsttimethatrecombinantASwithnosignalsequenceproducedinE.
colimostlylocalizesintheperiplasm.
TheC-terminal99-to-140sequenceisthemainpartresponsibleforthetranslocationofAS.
TheN-terminal1-to-60regionisnotrequiredfortranslocationofASintotheperiplasm.
PublishedinJ.
Bacteriol(2007).
2.
StudyonthecrystalstructureofERp44Asamemberofthehumanproteindisulfideisomerase(PDI)family,ERp44playsanimportantroleintheendoplasmicreticulum(ER)proteinqualitycontrol.
WesolvedthecrystalstructureofERp44at2.
6A,whichhasbeendepositedtoPDB(ID:2r2j).
ThisisthefirstfulllengthstructureofthehumanPDIfamilymembers.
AflexibleC-terminaltailturnsbacktotheb'andadomains,shieldingahydrophobicpocketindomainb'andahydrophobicpatchclosetothe-CRFS-motifindomaina.
RemovaloftheC-tailsignificantlyincreasestheenzymeandchaperoneactivities,andthefractionofERp44thatformsmixeddisulfideswithendogenousproteinsatsteadystate.
发表文章:G.
P.
Ren,X.
Wang,S.
F.
Hao,H.
Y.
Hu&C.
C.
Wang*.
Translocationofα-SynucleinexpressedinEscherichiacoli.
J.
Bacteriol.
2007;189,2777-2786.
28(三)生物膜与膜蛋白功能结构研究1、姬广聚组FK506结合蛋白对β-细胞钙释放及胰岛素分泌的影响研究及钙释放受体在发育期心肌胞的表达、功能及与先心病发生发展的可能关系均取得突破性进展;FKBP12.
6基因敲除对心肌细胞Ca2+释放受体功能的影响研究已成文;干细胞高效分化及应用研究取得阶段性进展.

1、Prof.
JiG.
J.
Duringtheyear2007alltheresearchprojectswecarryouthavebeenmadegreatprogress.
BesidesthefunctionalstudyonFKBP12.
6bindingproteinsinCardiac,smoothmyocytes,andpancreaticbeta-cells,themostexcitingthingsisthatwemakesignificantprogressinproteinERP44study.
WefindthattheERP44isnot,asreported,aIP3RsubtypeIselectiveinhibitor,butitpossessesalsomarkedlyeffectonRYRsinsmoothmuscle.
ThisencouragesustogenerateknockoutmousemodeltoinvestigatetheERP44indeeplyandbroadlyfurther.
AnotherexcitingeventinourresearchisthatwehavesuccessfullyconstructandexpressedspecificallytheG-CAMP2,auniqueCa2+indicator,incellnucleus.
ThisisveryimportantforustoinvestigatetheroleofnuclearinhandlingCa2+signalingforsomegeneticdiseases.
Additionally,wealsomakeremarkableprogressinourembryoheartstudy.
Wefindforfirsttimethata"switch"offunctionofRYRsandIP3Rsduringheartdevelopmentexsits.
292、焦仁杰组1)证明WRN是CAF-1行使功能所必须(Oncogene,2007).
2)证明dCAF-1-p180在染色质结构的建立与维持中至关重要(Dev.
Biol.
,2007).
目前正在研究该基因在表观遗传调控中的分子作用机制,已有一些很有意义的发现;同时我们也正在研究p180与p55的关系.
3)我们已得到smallboy基因的果蝇突变株.
该基因的纯合突变果蝇幼虫期致死,同龄幼虫个体比野生形小很多;而杂合突变果蝇寿命减短.
目前正在检测它的抗氧化能力变化、细胞生长及其与Insulin等信号通路的关系.
4)我们已获得果蝇dRecQ4和dRecQ5的突变株.
dRecQ4突变株对特异DNA损伤敏感,内复制(Endo-replication)减少,细胞周期发生变化,dRecQ5突变影响DNA双链断裂的修复能力,它可能直接参与同一染色体内同源重组的调控,而对染色体间的同源重组不重要.
5)合作项目包括:(a)Bcd调节基因表达的新机制;Branching基因与细胞生长的关系.
(b)dCAF-1等蛋白复合物的结构解析.
(c)dHDAC6与蛋白清除及Parkinson's病的关系.
我们已获得dHDAC6的LOF及GOS果蝇模型.
(d)5HTs与Octopamine受体(GPCRs)在睡眠、侵略性及抑郁症等行为方面的功能.
为此,我们正在制备6个受体突变株果蝇(已获得两个).
2、Prof.
JiaoR.
J.
TheJiaolabisfocusedontheresearchofepigeneticregulationofchromatinplasticityandgenomestabilityusingDrosophilaasamodelsystem.
Inthelasttwoyears,(1)WedemonstratedthatWRNisrequiredforCAF-1inresponsetoDNAdamage(2)dCAF-1-p180isessentialforDrosophiladevelopmentandmaintenanceofepigeneticmemory(3)LossoffunctionofdRecQ4affectsparticularDNAdamagesensitivity,endo-replicationandcellcyclewhiledRecQ5maybeessentialforhomologousrecombinationinthesamechromosome(4)smallboygenehasmultiplerolesincellgrowth,anti-oxidativestressandaging.
Inaddition,wearecollaboratingwithseverallabswithinChinaandtheworldtryingtounderstand(a)theroleofbranchinggene(b)thestructureofdCAF-1(c)theroleofdHDAC6intheprocessofproteinclearance(d)therolesofGPCRsinsleep,aggressionanddepressionetc.
发表文章:1.
Jiao,R.
*,J.
Harrigan,I.
Shevelev,T.
Dietschy,N.
Selak,F.
E.
Indig,J.
Piotrowski,P.
Janscak,V.
A.
BohrandI.
Stagljar*,2007.
TheWernersyndromeproteinisrequiredforrecruitmentofchromatinassemblyfactor1followingDNAdamage.
Oncogene,2007;26:3811-3822.
2.
Song,Y.
,He,F.
,Xie,G.
,Guo,X.
,Xu,Y.
,Chen,Y.
,Liang,X.
,Stagljar,I.
,Egli,D.
,Ma,J.
andJiao,R.
*,2007.
CAF-1isessentialforDrosophiladevelopmentandinvolvedinthemaintenanceofepigeneticmemory.
Dev.
Biol.
2007;311:213-222.
303、苗龙组本研究组以线虫精子为研究对象,研究细胞运动的调控机理.
结合运用Ascarissuum精子细胞的无细胞体系在生化上的可操作性以及C.
elegans在遗传学上的可操作性,研究在精子发生过程中的信号转导.
运用反向遗传学结合细胞生物学观察,发现在C.
elegans精子细胞活化及运动的过程中,小G蛋白和磷酸酶可能扮演重要角色,目前我们正在研究精子细胞活化过程中其信号转导通路.

我们也利用Ascarissuum精子的无细胞体系研究发现,其精子的活化过程与C.
elegans精子活化高度同源,来自C.
elegans的识别膜状体(MembranousOrganelle,MO)的抗体,也特异性地识别Ascarissuum精子细胞中的膜状体.
生化分析结合免疫荧光观察表明,该抗体标记的分子可能是一种膜结合蛋白,精子活化时,在膜状体与质膜融合的过程中,被释放到精子细胞表面.
在精卵识别的过程中,该分子可能起重要作用.
3、Prof.
MiaoL.
MygroupisstudyingtheamoeboidcellmotilityusingspermofnematodesAscarissuumandC.
elegans.
CombiningthebiochemicalaccessibilityofAscarissuumspermandgeneticsadvantagesofC.
elegans,wearenowstudyingthesignaltransductionduringthewormspermatogenesis(includingmeiosisandspermactivation).
Usingthegenomewidescreeningforinfertility,wefoundthatGTPasesandphosphatasesmayplaypivotalrolesduringC.
elegansspermiogenesisandspermmotility.
WealsofoundthatthespermactivationinAscarissuumisverysimilartothatinC.
elegans.
TheantibodyrecognizingtheMO(MembranousOrganelle)structureinC.
elegansalsolabeledtheMOsofAscarissuumspecifically.
FurtherbiochemicalandimmunofluorecencestudiessuggestedthattheantigenrecognizedbyMOantibodyisprobablyamembraneassociatedprotein.
DuringtheMOfusiontoplasmamembrane,itmaybereleasedtotheoutsideofplasmamembranetoactasanimportantsignalforsperm-eggrecognition.
发表文章:1、MiaoL,YiK,MackeyJM,RobertsTM.
ReconstitutioninvitroofMSP-basedfilopodiumextensioninnematodesperm.
CellMotilityandCytoskeleton.
2007;64:235-2472、苗龙.
细胞运动、细胞迁移与细胞骨架研究进展.
生物物理学报,2007;23:281-289.
314、徐涛组胰岛素在GLUT4转运过程中的关键作用位点的识别和确认胰岛素刺激下脂肪和肌肉细胞中的葡萄糖转运体4(GLUT4)会被大量转运到膜上,对维持血糖平衡起着很重要的作用.
一直以来,局限于传统的生化手段,GLUT4转运的具体步骤没能很好的区分和定义开来.
我们发展了单个GLUT4储存囊泡的标记和识别技术,在活细胞中实现了对其动态循环过程的跟踪.
依靠该技术我们确定了胰岛素调控GLUT4上膜的关键步骤是在囊泡锚定到细胞膜之后,主要是增强GLUT4储存囊泡与脂膜的融合能力.
我们还发现PI3K以及它的下游效应分子AS160参与调控囊泡的锚定步骤,初步揭示GLUT4储存囊泡与细胞膜锚定的分子机制.

应用模式生物线虫研究调控型分泌的分子机制模式生物线虫是很好的研究遗传和发育的系统,但其在细胞生物学特别是膜转运领域的贡献却十分有限.
主要的原因是缺少高时空分辨的功能研究手段.
我们克服了这个技术局限,首次将高时空分辨的分泌检测技术应用在线虫上,建立了在线虫细胞水平研究调控型分泌的技术平台.
利用该技术平台,我们证明了核心致密囊泡的胞吐过程需要一种称为UNC-31的蛋白,阐明了该蛋白参与囊泡锚定的作用机制,并发现了UNC-13(Munc13-1在线虫中的同源蛋白)和UNC-31蛋白存在相互作用.
该工作开辟了利用线虫模式生物研究囊泡分泌的新方向.
4、Prof.
XuT.
GLUT4-containingVesicleRanslocationinAdipocytesWedevelopedmethodtodissectdifferentstepsalongthetranslocationandfusionofGLUT4-containingvesiclesinliveadipocytes.
Usingthismethod,weidentifiedthekeystepofGLUT4vesicletranslocationthatisregulatedbyinsulin(Baietal.
,CellMetabolism,2007).
NovelInsightsofDenseCoreVesicleDockingfromStudiesonC.
elegansNeuronsWeadvancedtechniquesinrecordingexocytosisathightemporal-andspatial-resolutionfromC.
elegansneurons.
Usingthesetechniques,weinvestigatedtherequirementofUNC-31proteininthereleaseofdensecorevesiclesandrevealedanovelinteractionbetweenUNC-31andUNC-13inthedockingofdensecorevesicles.
ThisworkrepresentsanewhallmarkofusingC.
eleganstostudythebasicmechanismofregulatedexocytosis(Zhouetal.
,Neuron,2007).
发表文章:1)LiBai,YanWang,JunmeiFan,YuChen,WeiJi,AnlianQu,PingyongXu,DavidE.
James,andTaoXu.
DissectingmultiplestepsofGLUT4traffickingandidentifyingthesitesofinsulinaction.
CellMetabolism.
2007;5,47-57.
2)Ke-MingZhou,Yong-MingDong,QianGe,DanZhu,WeiZhou,Xian-GuangLin,TaoLiang,Zheng-XingWu,TaoXu.
PKAactivationbypassestherequirementforUNC-31inthedockingofdensecorevesiclesfromC.
elegansneurons.
Neuron.
2007;56:657-669.
3)ZhengzhengLi,JingzeLu,PingyongXu,XiangyangXie,LiangyiChen,andTaoXu.
MappingtheInteractingDomainsofSTIM1andOrai1inCa2+Release-activatedCa2+ChannelActivation.
TheJournalofBiologicalChemistry.
2007;282(40),29448–29456.
4)HuiLi,JingYao,XiaotianTong,ZhaohuaGuo,YingWu,LiangSun,NaPan,HoumingWu,TaoXu,32andJiupingDing.
InteractionSitesbetweentheSlo1PoreandtheNH2Terminusofthe_2Subunit,ProbedwithaThree-residueSensor.
TheJournalofBiologicalChemistry.
2007;282(24),17720–17728.
5)WEILI,SHANG-BANGGAO,CAI-XIALV,YINGWU,ZHAO-HUAGUO,JIU-PINGDING,ANDTAOXU.
CharacterizationofVoltage-andCa2+-ActivatedK+ChannelsinRatDorsalRootGanglionNeurons.
JournalofCellularPhysiology.
2007;212(2),348-357.
6)ZhuD,ZhouW,LiangT,YangF,ZhangRY,WuZX,XuT.
SynaptotagminIandIXfunctionredundantlyincontrollingfusionporeoflargedensecorevesicles.
BiochemBiophysResCommun.
2007;361(4):922-7.
7)CongMA,HaiHOU,WeiTIAN,andTaoXU.
Expression,PurificationandCharacterizationofCriticalDomainsofMunc13-1.
ActaBiochimicaetBiophysicaSinica.
2007;39(8):617-23.
335、杨福全组完善了高灵敏和高通量的多维蛋白质鉴定系统,在低丰度蛋白质、膜蛋白质以及蛋白质复合物鉴定中形成特色.
建立复杂生物样品目标蛋白质的分离、鉴定方法.
成功鉴定了疾病相关的潜在的蛋白质生物标志物.
利用SILAC策略研究L6成肌细胞分化前后的蛋白质表达水平的变化.
共定量分析了894个蛋白质.
其中267个蛋白质的表达水平上调,20个表达水平下调.
主要包括细胞信号转导、蛋白质生成与降解、物质代谢、分子伴侣、细胞黏附、细胞结构与运动、物质转运等相关蛋白.

利用SILAC策略研究胰岛素短时刺激下L6骨胳肌细胞从内膜系统向细胞膜转位的蛋白质动态变化.
定量分析胰岛素短时刺激下的L6骨胳肌细胞膜、低密度囊泡(LDM)组份中蛋白质组变化.
研究高糖状态下胰岛β细胞功能改变及其线粒体的蛋白质组学变化.
高糖处理后的INS-1细胞,其线粒体变圆、线粒体肿胀、线粒体比重增加、线粒体嵴减少、线粒体呼吸功能下降,INS-1细胞氧自由基生成增加,凋亡明显增加,ATP生成明显减少,胰岛素生成与分泌减少.
结果表明代谢相关蛋白、呼吸链相关蛋白、线粒体生成相关蛋白以及ROS相关蛋白发生了表达下调或上调.
MALDI-TOF-MS检测磷酸化肽段的技术方法的优化.
利用SILAC定量策略对胰岛素短时间刺激作用前后L6肌管细胞中可溶性磷酸化蛋白质组变化进行研究.
共鉴定到171个磷酸化蛋白,387个磷酸化位点,其中353个为未报道的磷酸化位点.
开展了脂质分析检测方法的初步研究.
建立了生物样本中脂质的分类提取、分离方法以及基于质谱的检测方法.
5、Prof.
YangF.
Q.
1)Optimizedthesensitiveandhighthrough-putmultidimensionalproteinidentificationtechniquesystemandwidelyusedfortheidentificationoflowabundantproteinandmembraneproteinsandproteincomplex.
2)Establishedmethodsforthepurificationandidentificationoftargetproteinsfromcomplexbiologicalsampleandsuccessfullyidentifiedseveralpotentialproteinbiomarkers.
3)PreliminaryquantitativeprofileofdifferentialexpressionbetweenratL6myoblastsandmyotubesbySILAC.
894proteinswerequantified.
267ofthemwereup-regulatedand20ofthemweredown-regulated.
Thesedifferentiallyexpressionalproteinsaremainlyinvolvedininter-orintracellularsignaling,proteinsynthesisanddegradation,molecularchaperone,celladhesionandextracelluarmatrix,cellstructureandmotility,metabolism,substancetransportation,etc.
4)SILACquantitativeproteomicmethodwasappliedtoanalyzeproteinsdynamicchangefromintracellularmembranecompartmentstoplasmamembraneofL6skeletalmusclecellsafterrapidinsulinstimulation.
5)Studyonproteomicchangesofmitochondriaofpancreaticβcellsaftertreatedbyhighglucose.
6)ThemorphologyandfunctionofmitochondriaofratderivedIns-1cellswerefoundalteredsignificantlyaftertreatedbyhighglucose.
Suchasroundprofiles,increaseddensity,decreasedridgesandincreasedbubblesofmitochondria,decreasedmembranepotentialandrespiratorycontrolrateofmitochondria,graduallyaccumulatedoxygenfreeradicalsinIns-1cells,increasedapoptosisofins-1cells,decreasedATPformationandinsulinsecretionofIns-1cells.
Thequantitativeproteomicresultindicatedthattheexpressionlevelofsomemitochondrialproteinssuchassubstratemetabolism,respiratorychain,mitochondriagenesis,apoptosisandROSformationrelatedproteins,changed.
Thestudypresentedmorecluesforimpairedinsulinsecretionormitochondriafunctionfailureindiabetic's34pancreaticβcells.
7)OptimizedthemethodfortheanalysisofphosphopeptidesbyMALDI-TOF-MS.
StudyonphosphoproteomicsofratL6myotubescellafterinsulinstimulationforshorttimebySILAC.
171phosphoproteinsand387phosphorylationsiteswereidentified.
Amongtheidentifiedphosphorylationsites,353werereportedforthefirsttime.
8)PrimarystudyontheanalysisandidentificationoflipidsbyMALDI-TOF-MSandLC-MS/MS.
356、杨福愉组系统研究了胰凝乳蛋白酶在细胞内的定位,并研究了其参与调控细胞凋亡的机制,发现胰凝乳蛋白酶定位在多种细胞的溶酶体中,并在细胞凋亡信号的刺激下从溶酶体中释放出来,其主要作用底物为Bid和Calcineurin;胰凝乳蛋白酶可将Bid剪切为tBid,tBid可快速诱导线粒体释放细胞色素c,并激活凋亡酶3依赖的细胞凋亡通路,启动细胞凋亡.
胰凝乳蛋白酶可将Calcineurin剪切为具有磷酸酶活性的大片段,且剪切后的Calcineurin其磷酸酶活性不再受钙信号调控.
研究了脂筏结构调控PMCA活性及PMCA与其他蛋白相互作用的机理,发现PMCA与nNOS存在部分共定位;在静息态细胞中,PMCA与nNOS不发生蛋白质相互作用;当细胞被钙信号激活后,PMCA与与nNOS发生蛋白质相互作用,,且该相互作用依赖于脂筏结构;转染外源PMCA后,细胞nNOS活性显著降低,NO生成受抑制,提示PMCA可去活化nNOS;PMCA对nNOS的去活化作用依赖于脂筏结构.
发现脂筏结构是调控典型纳米颗粒的跨膜运输的关键因素.
细胞主要通过clathrin-coatedpits运输富勒烯纳米颗粒;富勒烯纳米颗粒进入细胞后富集在溶酶体中,并可增强溶酶体的稳定性.

6、Prof.
YangF.
Q.
TheintracellularlocalizationoflysosomalchymotrypsinBwasinvestigated,anditsinvolvementinapoptosisregulationwasexplored.
ChymotrypsinBwerelocalizedinthelysosomesofvariouscelltypes,andcouldbereleasedintothecytosoluponapoptoticstimuli.
ItspotentialsubstratesincludeBidandCalcineurin.
Uponactivationbychymotrypsin-catalyzedcleavage,tBidcouldinducetherapidreleaseofmitochondrialcytochromec,whichactivatesthecaspase-30dependentapoptoticpathways,andinducesapoptosis.
Also,calcineurincouldbeactivatedbychymotrypsin-catalyzedcleavage.
TheregulationoflipidraftsonPMCAactivitywasinvestigated.
PMCAco-localizeswithnNOSinneuronalcells.
Insilentcells,PMCAshowsnoprotein-proteininteractionwithnNOS;uponcalciumactivation,PMCAinteractswithnNOSinalipid-raftdependentmanner.
ExogenousPMCAinactivatescellularnNOSandinhibitsthesynthesisofNO,suggestingthatPMCAisaregulatorofPMCA.
TheinactivationofnNOSbyPMCAdependsontheintegrityoflipidrafts.
Lipidraftplaysimportantrolesinthetransmembranetraffickingofnanoparticals.
Cellsuptakenanoparticalsviaaclathrin-coatedpitsmechanism.
Lysosomesaretheintracellularpoolfornanoparticals,andtheintegrityoflysosomalmembranecouldbestabilizedbynanoparticals.
发表文章:MelatoninimpairsNADPHoxidaseassemblyanddecreasessuperoxideanionproductioninmicrogliaexposedtoamyloid-1-42.
J.
PinealRes.
36(四)计算与系统生物学1、毕利军组DNA损伤是基因突变的诱因,导致基因组的不稳定性.
细胞依赖各种修复系统对其损伤进行修复.
已有证据显示人类DNA修复失活导致基因组DNA的高突变率,并引起多种癌症的发生,但具体的修复机制、各种修复途径、细胞周期调控及凋亡如何协同作用以保持基因组的稳定性和防止癌症的发生还远不清楚.
结核病是由结核杆菌引起的全球性传染疾病.
由于基因突变导致的耐药性使人类对其防治更加困难.
结核杆菌缺乏明显的错配修复基因、具有高突变率和易形成耐药性,目前还不了解这三个特征是否存在内在联系.
课题组围绕上述问题开展研究,主要工作概括为四个方面:1)DNA修复机制:完成了修复蛋白MutS寻找错配位点和UvrD的解螺旋功能可视化研究;研究修复蛋白MutL与DNA聚合酶III的相互作用揭示了DNA错配修复与复制系统的协同作用机制.
2)结核杆菌DNA修复与耐药性:完成了吡嗪酰胺酶的鉴定及其突变情况调查工作;开展GyraseB结构与功能研究;建立DNA修复蛋白库并开展DNA修复和耐药性的研究.
3)非编码RNA与DNA修复:垂钓ncRNA的靶蛋白并研究其功能;研究ncRNA对DNA修复功能的调控;开展结核杆菌ncRNA的标注.
4)分析生物技术:开展蛋白激酶检测肽芯片和基因突变检测等方法研究.
1、Prof.
BiL.
J.
DNArepairsystemtakesanimportantroleinkeepingstabilityofgenomeanditsdefectinthemammalianpathwayisassociatedwithastrongpredispositiontotumordevelopment.
Sofar,detailmechanismislessunderstood.
Tuberculosisisoneofthemostdeadlyandcommoninfectiousdiseases,whoseglobalspreadisfurthercomplicatedbytheubiquitousappearanceofdrug-resistantstrains.
OurworkhasbeenfocusedonthemechanismofDNAdamagerepairandtherelationshipbetweenDNArepairanddrug-resistance.
Furtherdetailsaregivenbelow:1)DNArepairsystem:(a)thesestudiesonthemechanismofMutSsearchingforamismatchsiteandVisualizationandkineticanalysisofDNAbindingandunwindingbyHelicaseII(UvrD)havebeendone.
(b)WefindthestrongevidencetosupportthenotionthatDNAreplicationandMMRarehighlyassociatedwitheachother.
2)DNArepairanddrug-resistanceofMTB:(a)CharacterizationofPncAandresearchontherelationshipofitsmutationanddrug-resistancehavebeenfinished.
(b)StudiesonstructureandfunctionofGyraseBhavebeendeveloped.
(c)WeareconstructingalibrarycomposedofDNArepairproteinsofMTBanddoingtherelativeworksonDNArepairanddrug-resistance.
3)Non-codingRNAandDNArepair:(a)wearebaitingthetargetproteinofncRNAintheC.
elegansandfurthertodiscovertheirfunctions.
(b)ResearchonregulationofncRNAonDNArepairfunctioninhumanisdoing.
(c)wearefocusingonfindingncRNAsanddiscoveringtheirfunctionsinmycobacteriumtuberculosis.
4)Analyticalmethods:theyinclu372、陈润生组研究组从事的非编码基因和系统生物学两个方面的研究工作都是国际的热点领域,本组又有多年的研究积累,所以进展顺利.
2007年已在国际SCI杂志上正式发表研究论文8篇,其中IF大于5的有4篇.
以通讯作者和生物物理所为第一单位发表的有4篇,其中包括:GenomeResearch一篇,NucleicAcidResearch一篇,BMCMolecularBiology一篇,PROTEINS.
一篇.
以署名作者发表的MolecularCell一篇等.
2、Prof.
ChenR.
S.
Ourgroupfocusontheresearchofnon-codingRNAandsystematicbiology.
Thetwoareasareboththeinternationalhotspots.
Meanwhile,wehaveaccumulatedmanyyearsofresearchexperiencesaboutthesefields.
Sotheworkisnowgoingsmoothly.
Intheyearof2007,wepublishedeightpiecesofresearchpapersontheSCImagazine.
TheaddressarealltheIInstitueofBiophysics.
TheImpactFactoroffourpiecesofthemaremornthan5.
Moreover,fourpiecesofpaperspublishedrespectievlyonGenomeResearch,NucleicAcidResearch,BMCMolecularBiologyandPROTEINS.
ThecoorespondentauthorofthesepapersisDr.
ChenandprimacyadressistheInstitueofBiophysics.
AlsoDr.
chenisthebylinerofonepaperinMolecularCell.
Thedetailisinthefollowingpaperlist.
发表文章:1.
HoushengHe,JieWang,TaoLiu,XShirleyLiu,TiantianLi,YunfeiWang,ZuweiQian,HaixiaZheng,XiaopengZhu,TaoWu,BaochenShi,WeiDeng,WeiZhou,GeirSkogerb,andRunshengChen,"MappingtheC.
elegansnon-codingtranscriptomewithawholegenometilingmicroarray",GenomeResearch.
2007;7,1-7.
2.
ShunminHe,ChangningLiu,GeirSkogerb,HaitaoZhao,JieWang,TaoLiu,BaoyanBai,YiZhaoandRunshengChen.
*NONCODEv2.
0:decodingthenon-coding,NucleicAcidsResearch.
2007;1–3.
3.
DongJia,LunCai,HoushengHe,GeirSkogerbo,TianTianLi,MuhammadNAUMANAftabandRunshengChen,Systematicidentificationofnon-codingRNA2,2,7-trimethylguanosinecapstructuresinCaenorhabditiselegans,BMCMolecularBiology.
2007;8:86.
4.
LarisaLitovchick,SubhashiniSadasivam,LaurenceFlorens,XiaopengZhu,SeleneK.
Swanson,SoundarapandianVelmurugan,RunshengChen,MichaelP.
Washburn,X.
ShirleyLiu,andJamesA.
DeCaprio,"EvolutionarilyConservedMultisubunitRBL2/p130andE2F4ProteinComplexRepressesHumanCellCycle-DependentGenesinQuiescence".
MolecularCell.
2007;26:539–5515.
YifeiYin*,YiZhao*,JieWang,ChangningLiu,ShuguangChen,RunshengChen,HaitaoZhao.
antiCODE:anaturalsense-antisensetranscriptsdatabase.
BMCBioinformatics.
2007;8:3196.
LeiLi,XiangfengWang,RajkumarSasidharan,ViktorStolc,WeiDeng,HangHe,JanKorbel,XueweiChen,WarapornTongprasit,PamelaRonald,RunshengChen,MarkGerstein,XingWangDeng*,GlobalIdentificationandCharacterizationofTranscriptionallyActiveRegionsintheRiceGenome.
PLoSONE.
2007;14;2(3):e2947.
XiangfengWang*,HangHe*,LeiLi,RunshengChen,XingWangDeng,SonggangLi,NMPP:auser-customizedNimbleGenmicroarraydataprocessingpipeline.
Bioinformatics.
2006;22(23):2955-7.
383、蒋太交组1.
利用流感的全基因组序列,我们建立了模拟流感演化的新模型.
这个模型对预测流感流行,疫苗制备和防治流感将会发挥重要作用.
而且,通过模型的分析,我们发现流感流行的新规律.
该文已发表于GenomeResearch上.
2.
与流感中心舒跃龙教授和MIT的陈建柱教授合作,通过计算机建模,我们发现导致03-04年人流感严重致病性的分子机制.
该文正在准备之中.
3、Prof.
JiangT.
J.
ComputationalanalysisofhumaninfluenzaevolutionanditsmolecularmechanismWeproposedanovelmodeltointerprettheevolutionarypatternsofhumaninfluenzabasedonitswholegenomesequence,anddiscoverednewevolutionarycharacteristicsofhumaninfluenzaviruses.
ThisworkhasbeenpublishedonGenomeResearch.
InacollaborativeworkwithProfessorYuelongShuatChinaCDCandProfessorChenJianzhuatMIT,weelucidatedthemolecularbasisthatunderpinstheinfluenzaoutbreakin03-04influenzaseason.
Thisworkisalsobeingpreparedforpublication.
发表论文:XJDu,ZWang,APWu,LSong,HYHang,TJJiang.
NetworksofNucleotideCo-occurrenceCaptureCharacteristicsofHumanInfluenzaEvolution.
GenomeRes.
394、饶子和组本课题组研究重点之一是重要病毒(如:SARS、MHV、IBV、HCoV等冠状病毒及其他与人类疾病相关病毒)和与人类重大疾病相关的蛋白质,开展三维结构与功能研究以及相关抑制剂药物研发.
本年度解析MHV-A59毒株非结构蛋白nsp4C端结构域、IBV主蛋白酶及其与N3抑制剂复合物以及nsp3的ADRP结构域的三维结构;解析诺罗病毒表面抗原P结构域、结核杆菌丙二酰辅酶A转酰基酶以及SIV中Mamu-A*01与两类主要表面抗原决定簇复合物的晶体结构;已获得N3、N24、N27、H6、H16、H23及H24这些抑制效果较好的抑制剂,为相关药物的开发提供了实验依据.
人体重要生物活性酶的结构与功能研究是另一重点研究方向.
解析了人源半胱氨酸双加氧酶、人源异戊烯焦磷酸异构酶、RSCUT、长链烷烃羟化酶LadA、类ParB蛋白核酶及其与底物或辅因子的复合物结构.
开展细胞周期、转录调控及信号转导相关蛋白的结构与功能研究.
解析了减数分裂纺锤体相关蛋白spindlin1、人源APPL1N端BAR-PH结构域、酵母DCN-1晶体结构.
2007年,在国际学术刊物上发表研究论文21篇,申请发明专利5项,获得国家发明专利2项.
4、Prof.
RaoZ.
H.
In2007,themainresearchfocusofourgroupinvolvedthree-dimensionalstructure,function,proteinengineeringandnoveldrugdesignstudiesoftargetproteinsrelatedtoimportantvirusesandhumandiseases.
Thestructure,functionandrelatedinhibitordesignstudiesofnon-structuralproteinsandassociatedreplication-transcriptionmachineryfromcoronaviruses,includingSARS-CoV,MHV,IBV,HCoV-HKU1.
WedeterminedthethreedimensionalstructureoftheC-terminalcytoplasmicdomainofMHVA59non-structuralprotein4(nsp4-C),IBVMproanditscomplexwithaninhibitorN3,ADRPdomainofnsp3,Pdomainofthenoroviruscapsid,Mamu-A*01incomplexwithtwoimmunodominantepitopesandMtMCAT.
Therewassignificantprogressonthediscoveryofwide-spectruminhibitors.
Oftheinhibitorswehavedesigned,N3,N24,N27,H6,H23andH24allshowstronginhibitioneffects,thusprovidingasolidbasisforthedevelopmentofeffectiveanti-coronavirusdrugs.
Anotherkeyfocusofourresearchisthestructuralandfunctionalstudyofhumanenzymeswithimportantbiologicalactivities.
Wedeterminedthethree-dimensionalstructuresofhumancysteinedioxygenase,humanIPPisomerase,RSCUT,long-chainalkanemonooxygenaseLadA,ParBnucleases,orincomplexeswiththeirsubstratesandco-factors.
Theresultsprovideastructuralbasisforfurtherinsightsintothecatalyticmechanismsofhumanenzymes.
Weseektocarryoutstructuralandfunctionalstudiesuponimportantproteinsincellcycle,transcriptionregulationandsignaltransduction.
Crystalstructuredeterminationofhumanspindlin1,humanAPPL1N-terminalBAR-PHdomainmotifandyeastDCN-1providesbiochemicalbasisforcellcycleregulationandotherfunctions.
Atotalof21researchpaperswerepublishedininternationaljournals,5inventionpatentswereappliedforand2wereacceptedasnationalpatents.
发表文章:1.
ShawN,ZhaoM,ChengC,XuH,SaarikettuJ,LiY,DaY,YaoZ,SilvennoinenO,YangJ*,LiuZJ*,WangBC&RaoZ.
Themultifunctionalhumanp100protein'hooks'methylatedligands.
NatStructMolBiol.
2007;14(8):779-7842.
ZhaoQ,QinL,JiangF,WuB,YueW,XuF,RongZ,YuanH,XieX,GaoY,BaiC,BartlamM,Pei40X*&RaoZ*.
Structureofhumanspindlin1:tandemtudor-likedomainsforcellcycleregulationJ.
Biol.
Chem.
.
2007;282(1):647-6563.
YeS,WuX,WeiL,TangD,SunP,BartlamM&RaoZ*.
Aninsightintothemechanismofhumancysteinedioxygenase:Keyrolesofthethioether-bondedtyrosine-cysteinecofactor.
JBiolChem.
2007;282(5):3391-34024.
YangX,ZhouJ,SunL,WeiZ,GaoJ,GongW,XuRM,RaoZ&LiuY*.
StructuralbasisforthefunctionofDCN-1inproteinNeddylation.
J.
Biol.
Chem.
2007;282(34):24490-244945.
XueX,YangH,ShenW,ZhaoQ,LiJ,YangK,ChenC,JinY,BartlamM&RaoZ*.
ProductionofAuthenticSARS-CoVMprowithEnhancedActivity:ApplicationasaNovelTag-cleavageEndopeptidaseforProteinOverproduction.
J.
Mol.
Biol.
.
2007;366(3):965-9756.
ZhengW,SunF,BartlamM,LiXM,LiR&RaoZ*.
Thecrystalstructureofhumanisopentenyldiphosphateisomeraseat1.
7Eresolutionrevealsitscatalyticmechanisminisoprenoidbiosynthesis.
J.
Mol.
Biol.
2007;366(5):1447-14587.
WuB,LiuY,ZhaoQ,LiaoS,ZhangJ,BartlamM,ChenW&RaoZ*.
CrystalStructureofRS21-C6,InvolvedinNucleosideTriphosphatePyrophosphohydrolysis.
JMolBiol.
2007;367(5):1405-14128.
LiZ,HuangY,GeJ,FanH,ZhouX,LiS,BartlamM,WangH&RaoZ*.
TheCrystalStructureofMCATfromMycobacteriumtuberculosisRevealsThreeNewCatalyticModels.
JMolBiol.
2007;371(4):1075-10839.
CaoS,LouZ,TanM,ChenY,LiuY,ZhangZ,ZhangXC,JiangX,LiX&RaoZ*.
StructuralBasisfortheRecognitionofBloodGroupTrisaccharidesbyNorovirus.
JVirol.
2007;81(11):5949-595710.
RaoZ.
HistoryofproteincrystallographyinChina.
PhilosTransRSocLondBBiolSci,2007;362(1482):1035-104211.
BartlamM,XuYY&RaoZ*.
StructuralproteomicsoftheSARScoronavirus:amodelresponsetoemerginginfectiousdiseases.
JStructFunctGenomics.
2007,8(2-3):85-9712.
HuoX,SuD,WangAJ,ZhaiYJ,XuJX,LiX,BartlamM*,SunF*&RaoZ,.
PreliminarymolecularcharacteristicandcrystallizationofmitochondrialrespiratoryComplexIIfromporcineheart.
FEBSJournal.
2007;274(6):1524-152913.
PangX,XuF*,BellSG,GuoD,WongLL*&RaoZ.
Purification,crystallizationandpreliminarycrystallographicanalysisofcytochromeP450203A1fromRhodopseudomonaspalustris.
ActaCryst.
SectF.
2007;63(4):342-34514.
PengY,XuF*,BellSG,WongLL*&RaoZ.
CrystallizationandpreliminaryX-raydiffractionstudiesofaferredoxinreductasefromRhodopseudomonaspalustrisCGA009.
ActaCrys.
F.
2007;63(5):422-42515.
ShawN,ChengC,TempelW,ChangJ,NgJ,WangXY,PerrettS,RoseJ,RaoZ,WangBC&LiuZJ*.
(NZ)CH.
.
.
OContactsAssistCrystallizationofaParB-likeNuclease.
BMCStructBiol.
2007;7:4616.
XuF,BellSG,RaoZ*&WongLL*.
Structure-activitycorrelationsinpentachlorobenzeneoxidationbyengineeredcytochromeP450cam.
ProteinEngDesSel.
2007;20(10):473-8017.
ShawN,TempelW,ChangJ,YangH,ChengC,NgJ,RoseJ,RaoZ,WangBC&LiuZJ*.
CrystalstructuresolutionofaParB-likenucleaseatatomicresolution.
Proteins.
2007;70(1):263-26718.
ZhuG,ChenJ,LiuJ,BrunzelleJS,HuangB,WakehamN,TerzyanS,LiX,RaoZ,LiG,ZhangXC*.
StructureoftheAPPL1BAR-PHdomainandcharacterizationofitsinteractionwithRab5.
EMBOJ.
2007;26(14):3484-3493415、孙飞组本课题组利用电子显微镜和X射线晶体的方法开展膜蛋白、病毒以及生物超分子复合体的结构研究.
研究方向为现代结构生物学,包括X射线结构生物学、冷冻电镜结构生物学和冷冻电镜断层成像的细胞结构学三个大的方面.
目前的研究内容包括:1.
线粒体呼吸链复合物II的电子传递机制;2.
线粒体呼吸链复合物I同源蛋白——大肠杆菌NDH-1的结构研究;3.
细菌脂蛋白合成通路上重要膜蛋白的三维结构研究;4.
古菌二型分子伴侣的冷冻电镜和晶体学研究;5.
线粒体呼吸链相关的解耦联蛋白UCP的三维结构研究;6.
帕金森氏病相关蛋白LRRK2的晶体结构研究(与许挚恒合作);7真核生物染色质重组蛋白因子的结构研究;8兔出血症病毒衣壳三维结构研究;9细胞内吞作用的再循环过程相关蛋白ACAP1的结构研究.
研究进展主要有:完成线粒体复合物II与两种抑制剂的复合物晶体结构解析;获得了细菌脂蛋白合成通路上两个重要膜蛋白的晶体;获得了细胞内吞作用的再循环过程重要蛋白ACAP1的晶体;与王志珍院士研究组合作,解析了内质网蛋白质质量控制系统中第一个人源全长ERp44蛋白的三维晶体结构,目前正在进行总结和文章写作;与范祖森研究组合作解析了人源颗粒酶M及其底物多肽的晶体结构,目前正在进行总结;利用电镜和X射线晶体学开展利用二型分子伴侣α和β的结构研究,成功利用负染方法完成了α分子外形的三维重构,观察到了同一种复合体内的两种分子构像,此外还获得了α分子和β分子的晶体,进一步的冷冻电镜工作和晶体学工作正在开展中.

5、Prof.
SunF.
ResearchProjectsWewillfocusonthestructureofmembraneprotein,virusandbiologicalsupermolecularcomplexviaelectronmicroscopeandX-raycrystallographycombination,includingX-raystructuralbiology,cryo-electronmicroscopystructuralbiologyandcryo-electrontomographycellbiology.
Wehavealreadybegunseveralstructuralresearchprojects:1.
MitochondrialrespiratoryComplexIIstructuralelectrontransfermechanism;2.
StructuralstudyofmitochondrialrespiratoryComplexIhomology–E.
coliNDH-1;3.
3Dresearchofimportantbacteriallipoproteinsynthesispathwayrelatedmembraneproteins;4.
3Dresearchofsecondtypeofchaperonfromarchaeal;5.
StructuralresearchonmitochondrialrespiratoryrelateduncouplingproteinUCP.
6.
CrystalstructuralresearchonParkinsondiseasesrelatedproteinLRRK2(collaboratedwithZhihengXu).
7.
StructuralstudyofchromatinassemblyfactorcomplexinDrosophila8.
Structuralstudyofrabbithemorrhagicdiseasevirus(RHDV);9StructuralresearchonACAP1involvedinendocyticrecycling.
ResearchProgressWehavealreadysuccessfullysolvedtwocrystalstructuresofmitochondrialrespiratoryComplexIIwithtwoinhibitors.
Weobtainedthecrystalsoftwoimportantmembraneproteinsrelatingwithbacteriallipoproteinsynthesispathway.
AndwehaveobtainedthecrystalofACAP1.
Besides,withcollaborationfromProf.
WangC.
C.
,wesolvedthefirstcrystalstructureofhumanERp44relatingwithproteinqualitycontrolsysteminendoplasmicreticulum.
WehadthecollaborationwithProf.
FanZ.
andsolvedthecrystalstructureofhumanGzmManditscomplexstructureboundtopeptide.
Finally,wesuccessfullyreconstructedthe3DstructureofarchaealtypeIIchaperonαbynegativestainmethod,revealingtwodifferentconfirmationsinonecomplex,andthenwealsoobtainedcrystalsofchaperonαandchaperonβ;FurtherworkoncryoEMandX-raycrystallographyisnowgoing.
发表文章:1.
Huo,X.
,Su,D.
,Wang,A.
,Zhai,Y.
,Xu,J.
,Li,X.
,Bartlam,M.
*,Sun,F.
*,Rao,Z.
PreliminarymolecularcharacteristicandcrystallizationofmitochondrialrespiratoryComplexIIfromporcineheart,FEBSJ.
2007;274:1524-29.
2.
ZhengW,SunF,BartlamM,LiXM,LiR&RaoZ.
Thecrystalstructureofhumanisopentenyldiphosphateisomeraseat1.
7Aresolutionrevealsitscatalyticmechanisminisoprenoidbiosynthesis.
JMolBiol.
2007,366(5):1447-58.
42(五)感染与免疫的分子基础研究1、邓红雨组1)以小鼠疱疹病毒-68(MHV-68)为肿瘤相关疱疹病毒的模型,本课题组从以下几个方面对该病毒的复制机理展开了研究:(1)基因表达的转录调控及表观遗传学:病毒编码的复制和转录因子RTA在MHV-68生命周期中起着分子开关的重要作用.
我们的研究表明,rta基因启动子上组蛋白的乙酰化能激活MHV-68从潜伏期进入裂解期.
同时我们还对rta基因自激活及受病毒颗粒蛋白激活的具体调控机理进行了研究.
(2)基因组复制:鉴定了MHV-68基因组的左端复制子,并详细研究了相关的顺式元件.
首次发现细胞转录因子NF-Y在病毒基因组复制中起重要作用,并对其作用机理展开了详细研究.
(3)病毒包装:结合遗传学,蛋白质组学,结构学及电子显微镜技术选择性地对MHV-68的间质蛋白ORF33和ORF52在病毒颗粒的组装和出胞过程中的作用和机理进行了研究.
2)以鼠肝炎病毒(MHV-A59)为急性感染和免疫的模型,通过靶向重组,构建了携带T细胞表面抗原的重组病毒,研究病毒感染后小鼠T细胞的各种反应.
3)通过构建鼠肝炎病毒/人丙肝病毒嵌合病毒,研究丙肝病毒慢性感染的机制.

1、Prof.
DengH.
Y.
Weinvestigatethereplicationmechanismsofmurineherpesvirus-68(MHV-68)asamodelfortumor-associatedherpesviruses.
Specifically,wehavecarriedoutresearchinthefollowing3directions:1)TranscriptionalandepigeneticregulationofMHV-68geneexpression.
WehavefocusedonregulationoftheRTAgene,whoseproductisthe"molecularswitch"forMHV-68lifecycle.
WehaveshownthathistoneacetylationplaysanimportantroleinmediatingreactivationofMHV-68fromlatency.
WearealsodissectingthemechanismsgoverningactivationofthertapromoterbyvirioncomponentsaswellasRTAproteinitself.
2)Viralgenomereplication.
Wehaveidentifieda1.
1-kboriginoflyticreplication(oriLyt)locatedtowardtheleftviralgenome.
Throughasystematicapproach,wehaveidentifiedthecriticalcis-elementsinvolvedinmediatingviralDNAreplication.
WehaveshownthatacellulartranscriptionfactorNF-Yplaysanimportantroleinthisprocess.
Toourknowledge,thisisthefirsttimethatNF-YhasbeenimplicatedinmediatingDNAreplication.
Wearefurtherinvestigatingthedetailedmechanisms.
3)Virionmorphogenesisandegress.
Wecombinegenetics,proteomics,structuralbiologyandelectronmicroscopyapproachestostudyviraltegumentproteins(currentlyfocusingonORF33andORF52)andtheirroleinvirionpackagingandegress.
Weemploymurinehepatitisvirus-A59(MHV-A59)asamodelsystemtostudyacuteviralinfectionandimmunity.
Throughtargetedrecombination,wehavegeneratedarecombinantMHV-A59thatcarriesT-cellepitopestostudyTcellresponsestocoronavirusinfectioninvivo.
Weinvestigatethemechanismsresponsibleforchronic/persistentinfectionofHCVthroughconstructingchimericMHV-A59/HCVvirusesandstudyingtheimmuneresponsesinvivo.
发表文章:431.
Deng,H.
,Y.
Liang,andR.
Sun.
RegulationofKSHVlyticgeneexpression.
CurrentTopicsinMicrobiologyandImmunology,2007;312:157-83,2.
YuF.
,JNHarada,HJBrown,H.
Deng,MJSong,TTWu,J.
Kato-Stankiewicz,CGNelson,J.
Vieira,F.
Tamanoi,SKChanda,andR.
Sun.
SystematicidentificationofcellularsignalsreactivatingKaposi'sarcoma-associatedherpesvirus.
PLoSPathogens2007;3(3):e443.
WongE.
,TTWu,N.
Reyes,H.
Deng,andR.
Sun.
Murinegammaherpesvirus68openreadingframe24isrequiredforlategeneexpressionafterDNAreplication.
JournalofVirology2007;81:6761-6764,.
4.
BortzE.
,L.
Wang,Q.
Jia,TTWu,JPWhitelegge,H.
Deng,ZHZhou,R.
Sun.
MurineGammaherpesvirus-68ORF52EncodesaTegumentProteinRequiredforVirionMorphogenesisintheCytoplasm.
JournalofVirology.
2007;81:10137-10150,.
5.
J.
Benach,L.
Wang,Y.
Chen,CKHo,S.
Lee,J.
Seetharaman,R.
Xiao,TBActon,GTMontelione,H.
Deng,R.
Sun,andL.
Tong.
StructuralandfunctionalstudiesoftheabundanttegumentproteinORF52frommurinegammaherpesvirus-68.
JournalofBiologicalChemistry2007;282:31534-31541442、范祖森组阐明了颗粒酶M介导靶细胞凋亡机制在发现颗粒酶M引起DNA双链断裂为特征的靶细胞凋亡的基础上(Journalofimmunology,2006).
我们继续研究发现颗粒酶M进入靶细胞后直接作用线粒体,从而引起活性氧的产生.
热休克蛋白75(HSP75)具有抑制活性氧产生的功能.
研究发现颗粒酶M进入靶细胞后,随即切割HSP75蛋白并破坏了其相应的抗氧化功能,打破了细胞氧化平衡体系,从而决定细胞走向凋亡的命运.

该研究成果刚被JBC杂志接受发表.
我们表达了具有高度活性的颗粒酶M,获取了其大量均一的表达.
与孙飞课题组合作通过优化晶体生长条件,获取了颗粒酶M的晶体.
收取了高分辨率的衍射数据,解析了颗粒酶M的结构.
正在探索结构与功能的关系.
揭示了颗粒酶H在NK/CTL介导肿瘤杀伤中的作用我们研究发现颗粒酶H进入肿瘤细胞后介导了快速的靶细胞凋亡.
磷脂酰丝氨酸外翻、染色体凝集、核形态发生变化及DNA双链断裂为颗粒酶H介导靶细胞死亡的主要特征.
颗粒酶H介导的靶细胞死亡依赖于Caspase的激活,以及CAD活性的释放.
并发现颗粒酶H能水解Bid为tBid,引起线粒体肿胀和破坏,引起cytochromec的产生,致使细胞走向凋亡.
我们正在从结构和功能的基础上,深入探明颗粒酶H介导肿瘤杀伤中的作用底物和机理.
2、Prof.
FanZ.
S.
GetinsightintothemechanismsofgranzymeM-mediatedtargetcellapoptosisWerecentlydemonstratedthatGzmMinducescaspase-dependentapoptosiswithDNAfragmentationthroughdirectcleavageofinhibitorofcaspase-activatedDNase(ICAD).
However,themolecularmechanismsforGzmM-inducedapoptosisisunclear.
WefoundGzmMcausesmitochondrialswellingandlossofmitochondrialtransmembranepotential.
MoreoverGzmMinitiatesreactiveoxygenspecies(ROS)generation.
Heatshockprotein75(HSP75)actsasanantagonistofROSandprotectscellsfromGzmM-mediatedapoptosis.
GzmMcleavesTRAP1andabolishesitsantagonisticfunctiontoROSresultinginROSaccumulation.
ROSaccumulationisinaccordancewiththereleaseofcytochromecfrommitochondriaandenhancesGzmM-mediatedapoptosis.
WeobtaineditscrystalofgranzymeMandanalyzeditsstructureincollaborationwithSun'sLab.
WeareinvestigatingthefunctionsofgranzymeMbasedonitsstructure.
ThefunctionsofgranzymeHinNK/CTL-mediatedcelldeathGranzymeH(GzmH)playsapivotalroleinNKcellmediatedcytolysis.
HoweverGzmHisdefinedasanorphangranzymeanditsfunctionhaslessbeendefined.
HerewedemonstrateGzmHcaninducerapidapoptosisoftargetcells,whichisdependentoncaspaseactivationandmitochondrialdamage.
GzmH-induceddeathischaracterizedbyphophatidylserineexternalization,nuclearcondensation,DNAfragmentation,caspaseactivationandcytochromecreleasethatarehallmarksoftypicalapoptosis.
GzmHcandirectlycleaveICADtounleashCADforDNAfragmentation.
Moreover,GzmHdirectlyprocessesBidtoproducetheactiveformtBidleadingtocytochromecrelease.
WearefurtherinvestigatingitsmechanismsofgranzymeH-mediatedtargetcellapoptosis.
发表文章:1.
ZhaoT,ZhangH,GuoY,ZhangQ,LuH,HouQ,HuaG,FanZ*.
GranzymeKInducesrapidcaspase-independentcelldeathwithapoptoticnuclearmorphologyandsingle-strandedDNAnicks.
CellDeathandDifferentiation2007;14:489-499.
2.
HuaG,ZhangQ,FanZ*.
Heatshockprotein75(TRAP1)antagonizesreactiveoxygenspeciesgenerationandprotectscellsfromgranzymeM-mediatedapoptosis.
JBiolChem.
2007;282(28):20553-60.
3.
ZhaoT,ZhangH,GuoY,FanZ*.
GranzymeKdirectlyprocessesBidtoreleasecytochromecandendonucleaseGleadingtomitochondria-dependentcelldeath.
JBiolChem.
2007;282(16):12104-11453、高光侠组本课题组主要研究以HIV为代表的逆转录病毒与宿主相互作用的分子机理,锌指结构抗病毒蛋白(ZAP)的作用机理是本实验室的研究重点.
我们前期的工作发现ZAP通过特异性地降低细胞质中病毒mRNA的稳定性从而抑制病毒的复制.
ZAP本身不具有RNA酶的活性,但具有反式作用因子的功能,能够直接结合特定的病毒RNA,并招募RNA降解机器实现对靶RNA的降解.
本年度取得主要成果如下:1)分析了RNA降解机器Decapping和Deadenylation复合物与ZAP之间的相互作用关系,发现ZAP与PARN有直接的相互作用,用RNAi方法敲低PARN可明显减弱ZAP的活性.
同时发现,5'-3'外切酶XrnI参与ZAP介导的RNA降解.
2)改进了鉴定ZAP复合物组分的方法,鉴定出多个可能参与调控ZAP功能的细胞因子.
根据作用机理,大致可将其分为三类.
第一类是为ZAP实现其功能所必需的,如RNA解旋酶p72;第二类参与调控ZAP的控能,如TRIM25;第三类通过结合ZAP作用的靶RNA拮抗ZAP的功能,如YB1.
3)利用RNAi方法敲低p72、TRIM25都可明显减弱ZAP的活性;过表达YB1也会明显减弱ZAP的活性.
上述结果表明这些细胞因子在ZAP功能中发挥着重要作用,具体的作用机理正在研究中.
3、Prof.
GaoG.
X.
Ourresearchmainlyfocusesonthemolecularmechanismoftheinteractionbetweenretrovirusesandtheirhosts.
Specifically,weworkonthemechanismsofthehostantiviralfactorZAP,whichinhibitsvirusinfectionbyspecificallydegradingviralmRNAs.
OurpreviousstudiesrevealthatZAPspecificallyeliminatestheviralmRNAinthecytoplasm.
OurcurrentworkingmodelisthatZAPdirectlybindstothetargetviralmRNAthroughthezinc-fingermotifs,andrecruitsthecellularRNAdegradationmachineries.
AnalysesoftheinvolvementoftheRNAdegradationmachineriesrevealedthatZAPrecruitsdeadenylasePARNtoshortenthepolyAtail.
The5'-3'exoribonucleaseXrnIisalsoinvolvedinZAP-mediatedmRNAdegradation.
InadditiontostudyingtheinteractionofZAPwiththeknowncomponentsoftheRNAdegradationmachineries,weusedvariousmethodstoidentifyproteinsassociatedwithZAP.
Ahandfulofcandidateproteinshavebeenidentified,severalofwhichhavebeenconfirmedtobeinvolvedinZAP-mediatedRNAdegradation.
BasedonthemechanismsbywhichtheseproteinsaffectZAP'sactivity,theycanberoughlyclassifiedintothreegroups.
TheproteinsingroupI,suchasTRIM25andGSK3,modulateZAP'sactivitythroughbindingtoormodificationofZAP.
TheproteinsingroupII,suchasYB-1andIMP1,seemtoantagonizeZAP'sactivitythroughbindingtoZAP'stargetRNAandtherebypreventtheRNAfromZAP-mediatedRNAdegradation.
TheproteinsingroupIII,suchasDEADboxRNAhelicasep72,functionascofactorsofZAP.
ThecurrentworkingmodelisthatZAPbindstothetargetRNAthroughthezinc-fingermotifs,recruitstheRNAhelicasep72tounwindtheRNAandrecruitstheRNAdegradationmachineriestodegradetheRNA.
DetailedmechanismsoftheseproteinsinZAP-mediatedRNAdegradationarecurrentlyunderinvestigation.
发表文章:GuoX.
,MaJ.
,SunJ.
,andGaoG*.
TheZinc-fingerAntiviralProteinRecruitstheRNAprocessingExosometoDegradeTargetRNAs.
ProcNatlAcadSciUSA.
2007;Vol.
104(1):151-6464、秦志海组本研究组主要运用转基因动物研究细胞因子对肿瘤间质细胞的作用及其机制,研究发现细胞因子介导的抑制血管新生是重要的抗肿瘤免疫效应机制.
主要进展如下:1)天然免疫细胞是肿瘤排斥过程中IFN-γ的重要产生者,而T细胞起辅助作用.
我们运用IFN-γ-/-及Rag1-/-小鼠构建了只有天然免疫细胞才具有IFN-γ产生能力的骨髓嵌合小鼠.
研究显示T细胞并不是必不可少的IFN-γ产生者,天然免疫细胞可以产生足够量的IFN-γ,而且其产生IFN-γ的能力只有在T细胞存在的情况下才能激活.
T细胞和天然免疫细胞之间的相互作用可能由IL-2介导.
为了进一步研究IFN-γ在抑制血管新生中的重要作用,我们还成功构建了组织特异性IFN-γ受体转基因小鼠,这一模型将有助于进一步阐明IFN-γ的作用机制.
2)肿瘤坏死因子TNF-α通过巨噬细胞来源的一氧化氮抑制肿瘤内血管形成.
肿瘤坏死因子(TNF)有两个受体,但是关于TNFR2的作用研究甚少.
本组研究首次证明TNFR2在机体天然免疫细胞上的表达可以独立介导TNF的抗肿瘤作用,而NO及其对肿瘤血管新生的抑制是这一过程所必需的.
4、Prof.
QinZ.
H.
Usingtransgenicanimals,wehaveinvestigatedtheeffectsofcytokinesontumorstromalcellsandtheunderlyingmechanisms.
Ourresultsindicatethatcytokinemediatedangiostasisisthemajorimmunologicalmechanismfortumorrejection.
Wefound:1)InnateimmunecellsarethemajorproducerofIFN-γ,andTcellscooperatewithinnateimmunecellsforIFN-γ-mediatedtumorrejection.
Byusingvariousbonemarrowchimericmice,weshowherethatIFN-γessentialfortumorimmunityissolelyproducedbyhemopoieticcells.
Surprisingly,IFN-γderivedfromTcellswasnotnecessaryfortumorimmunityinthismodel.
Intheimmunizedmice,inwhichonlyinnateimmunecellshavetheIFN-γ-producingpotential,tumorswereefficientlyrejected.
Theinnateimmunecells,suchasNK1.
1+cellsandCD11b+cells,canprovidesufficientamountsofIFN-γwhichrequires,however,thehelpofTcells.
TheclosecooperationbetweenTcellsandinnateimmunecellsduringtumorregressionislikelymediatedbyIL-2.
2)Tumornecrosisfactorinhibitstumorangiogenesisviamacrophages-derivednitricoxide.
Tumornecrosisfactor(TNF)bindstotwodifferentreceptors.
AlthoughmostofitsfunctionsareattributedtoTNFreceptor1(TNFR1),theindependentroleofTNFR2isstilllargelyunknown.
OurresultsshowforthefirsttimethatTNFR2expressedonhostinnateimmunecellsissufficienttomediatetheantitumoreffectofTNF,andNOisnecessaryforthisprocess,possiblybyinhibitionofangiogenesisinthetumor.
发表文章:1.
Zhao,X.
,M.
Mohaupt,J.
Jiang,S.
Liu,B.
Li,andZ.
Qin.
TumorNecrosisFactorReceptor-2-MediatedTumorSuppressionisNitricOxideDependentandInvolvesAngiostasis.
CancerRes.
2007;67(9):4443-50.
2.
Li,Z.
,P.
Felicia,B.
Li,S.
Liu,andZ.
Qin.
CrosstalkbetweenTcellsandinnateimmunecellsiscrucialforIFNγ-dependenttumorrejection.
JImmunol.
2007;179(3):1568-76.
475、唐宏组研究炎症反应中的信号转导机制,细胞凋亡机理,病毒感染的天然免疫调控机理.
2006-2007年的研究工作已发表高水平论文多篇.
5、Prof.
TangH.
Majorresearchinterestsfocusonsignaltransductionmechanismininflammationreaction,apotosismechanism,virusinfectionandinnateimmununeregulation.
发表文章:1.
JianjunChen,LuChen,GangWang,HongTang,Cholesterol-Dependentand-IndependentCD40InternalizationandSignalingActivationinCardiovascularEndothelialCells,ArteriosclerThrombVascBiol.
2007;27:2005-20132.
Yang-xinFu,kwangdongKim,JieZhaoandHongTang,Adaptiveimmunecellstemperinitialinnateresponses,NatureMedicine.
2007;13:1248-12523.
Leipan,ShuyiChen,ChangjiangWeng,Geraldcall,DengxiaoZhu,HongTangandTingXie,Stemcellagingiscontrolledbothintrinsicallyandextrinsicallyinthedrosophilaovary,CellStemCell.
2007;1:458-469486、唐捷组目前本课题组在下面三个方面取得阶段性进展:1)发现MIF在LPS诱导的iNOS上调中的关键作用.
同时发现MIF内吞对其信号转导具有关键作用,MIF可以直接进入细胞核,影响Jab1的细胞定位.
2)发现CLM家族蛋白在LPS诱导的TNF-alpha分泌中的作用.
3)发现在类风湿关节炎病人中清除B细胞后,引起T细胞和巨噬细胞功能的改变.
6、Prof.
TangJ.
Wemadeprogressinthefollowingaspects:1)MIFfunctionisimportantforLPSinducediNOSup-regulation.
MIFendocytosisisessentialforitssignaltransductionanditentersnucleustointeractwithJab1.
2)CLMfamilyreceptorsareinvolvedinLPSinducedTNF-alphasecretion3)BcelldepletioninrheumatoidarthritispatientsledtoTcellandmacrophagefunctionchanges.
发表文章:1.
TheHMGB1acidictailregulatesHMGB1DNAbindingspecificitybyauniquemechanism.
Biochem.
Biophy.
Res.
Comm.
2007;360(1):14-92.
IdentificationofeightnovelalternativesplicingformsofCD72andtheirdifferentialexpressioninamousemodelofSLE.
Prog.
Biochem.
Biophys.
2007;34(11):1175-11813.
MIFregulatesTNFAlphaReceptorTypeIIExpressioninRAW264.
7Cells.
Prog.
Biochem.
Biophys.
2007;34(6):580-584497、王盛典组实验室的研究工作主要集中在免疫调节分子,主要是共刺激分子在慢性病毒感染、肿瘤等疾病中的免疫调节作用.
其中与他人合作研究发现,慢性乙肝病人mDC上的B7-H1分子表达增高,可抑制抗HBV的T细胞免疫反应;抗共刺激分子B7-H1、CTLA-4抗体可增强肿瘤疫苗的抗肿瘤效果,这些成果分别发表在JImmunol(2007)(共同通讯作者);Immunol.
Lett(2007)(共同通讯作者)和J.
Clin.
Immunol(2007).
通过基础研究与临床资源和需求的紧密结合,我们研究发现4-1BB共刺激信号途径在慢性乙肝的发生和发展中起重要作用;CD24基因的单核苷酸多态性与慢性乙肝病人的肝硬化和肝癌发生有相关性,动物实验结果也显示CD24分子可促进HBV转基因小鼠肝癌的发生.
此外,发现PD-1、B7-H3分子在慢性乙肝免疫耐受中有重要调控作用.
通过与协和肿瘤医院的密切合作,我们已对20多例结肠癌组织和前哨淋巴结中,重要免疫细胞和分子的功能进行了全面研究,结合体外及动物实验,初步获得了有关B7-H1/PD-1信号途径、Treg和MSC细胞在肿瘤免疫耐受中作用的重要发现.
经过一年多的努力,终于从美国合作实验室获得了RagγCDKO免疫缺陷小鼠,目前正通过协和动物所从国外运送,下一年的另一项重点工作是利用该小鼠,建立人源化的慢性乙肝和肿瘤模型,这一模型的建立将显著提升实验室的科研水平.
7、Prof.
WangS.
D.
Ourstudiesfocusontheregulatoryfunctionsofmolecules,especiallyco-signalingmoleculesintheimmuneresponsesagainstchronicvirusinfectionsandtumors.
WefoundincollaborationwithothergroupsthattheexpressionofB7-H1onmDCofPBMCisincreasedinpatientswithchronicHBVinfectioncomparedwithhealthycontrols,andtheB7-H1expressedonmDCinhibitTcell-mediatedresponseagainstHBVinfection.
MonoclonalantibodiesagainstB7-H1orCTLA-4enhancetheanti-tumoreffectsoftumorvaccine.
TheseresultswerepublishedinJImmunol(2007)(co-respondingauthor);Immunol.
Lett(2007)(co-respondingauthor)andJ.
Clin.
Immunol(2007).
Combinedourbasicstudywithclinicalresourceandrequirement,ourstudiesshowedthatco-signalingof4-1BBplayaimportantroleinincidenceanddevelopmentofchronichepatitisB;ThesinglenucleotidepolymorphismsofCD24isrelatedwithdevelopingofcirrhosisandhepatocellularcarcinomainpatientswithchronicHBVinfection,andCD24moleculecanpromotethedevelopmentoflivercancerinHBVtransgenicmice.
Inaddition,wefoundthatco-signalingmoleculesofPD-1andB7-H3mayhaveanimportantrolesinregulationofimmunetoleranceinchronichepatitisB.
CollaboratingwithCancerInstituteaffiliatedwithPekingUnionMedicalCollege,westudiedthefunctionsofimportantlymphocytesandmoleculesintumortissuesanddraininglymphnodesfrommorethan20colorectalcancerpatientsandhassomeinterestingdiscoveriesaboutthefunctionsofB7-H1/PD-1co-signalingpathway,TregandMSCsintumor-inducedimmunetolerance.
AtlastwegotpermissionforRagγCDKOimmunedeficientmicefromoneofourcollaboratinglabsinUSthroughoneyearofhardwork.
OneoftheimportantworksfornextyearistoestablishthemodesofhumanizedmicewithchronichepatitisBandtumors,whichisveryimportantfordramaticallyraisingthelevelsofscientificresearchinourlab.
50发表文章:1.
LiN,LiN,QinH,LiX,ZhouC,WangD,MaW,LinC,ZhangY,WangS*,ZhangS*.
Synergisticantitumoreffectofchemotactic-prostatetumor-associatedantigengene-modifiedtumorcellvaccineandanti-CTLA-4mAbinmurinetumormodel.
ImmunolLett.
2007;113(2):90-98.
2.
ChenL,ZhangZ,ChenW,ZhangZ,LiY,ShiM,ZhangJ,ChenL,WangS*,WangFS*.
B7-H1upregulationonmyeloiddendriticcellssignificantlysuppressesT-cellimmunefunctioninpatientswithchronichepatitisB.
J.
Immunol.
2007;178(10):6634-41.
3.
LiN,QinH,LiX,ZhouC,WangD,MaW,LinC,ZhangY,WangS,ZhangS.
Potentsystemicantitumorimmunityinducedbyvaccinationwithchemotactic-prostatetumorassociatedantigengene-modifiedtumorcellandblockadeofB7-H1.
J.
Clinimmunol.
2007;27(1):117-130.
518、阎锡蕴组肿瘤新靶点及新功能抗体作用机理:(1)继续深入研究CD146分子及其抗体抑瘤作用机制,在活细胞上观察CD146二聚体与肿瘤化的相关性(BBA-molecularcellresearch1773:513,2007);(2)用Confocal观察到CD146分子具有类似细胞膜受体介导内吞的内在化现象;(3)继续寻找CD146天然配体,发现CD146的胞内区通过moesin的介导,与actin和vimentin等形成复合体,将相关信号传递下去;(4)构建抗CD146的人源化抗体,开发肿瘤抗体药物.
新型免疫检测方法:把肿瘤靶分子或抗体与纳米材料结合创造新型免疫磁珠.
在国际上首次报道磁性纳米颗粒具有类似过氧化物酶活性,提出纳米材料模拟酶的新概念,并利用其新特征建立一种新型免疫检测方法.
研究结果发表在NatureNanotechnology,同期杂志《新闻与观点》栏目配发了评论文章,称"这一发现不仅为惰性金属材料在纳米尺度具有催化活性的学说提出了新的证据,而且拓展了磁性纳米材料的应用".
随后美国Science杂志(ScienceNews,172:174,2007),美国Nanowerk(Nanowerk,Oct.
18th,2007)发表评述文章.
8、Prof.
YanX.
Y.
Themechanismstudyofanti-tumorantibody:Wehaveestablishedadifferentialscreeningplatformforspecifictumor-targetingantibodies;anddiscoveredseveraltumorbiomarkers.
Importantly,wehavefoundthatCD146isanoveltargetontumorbloodvesselsandinvolvedintumorangiogenesis.
ThefunctionandmechanismofCD146anditsantibodyAA98havebeenextensivelystudied:(1)WefoundthatCD146dimerizationwasup-regulatedbytumorconditionalmediumthroughtheNF-kappaBpathwayanddown-regulatedbymAbAA98.
(BBA-molecularcellresearch1773:513,2007).
(2)WeobservedtheendocytosisofCD146byusingTIRFM(Single-molecule)inlivingcells,suggestingtheregulabilityofthequantityofCD146onplasmamembraneanditssignificanceinregulatingsignaltransduction.
(3)WefoundthatCD146interactedwithactinandvimentinviamoesin,whichmightbeanimportantpathwaytotransferthesignaldownwards.
(4)WehaveconstructedthehumanizedCD146antibody,andtheinventionhasbeenlicensedbyanindustrialpartnerfordevelopingantibody-basedtumordrug.
Antibody-basedbiosensor.
Weconstructnovel-structuresofantibodycombinedwithnano-materialandnano-technologytomakenewbiosensorfordiagnosis.
Duringrunningthisproject,wefoundthatmagneticnanoparticles(MNPs)haveaperoxidase-likeactivity.
Basedonthisfinding,wehavedevelopedanovelimmunoassayusingthemulti-functionofantibody-modifiedMNPs(targeting,separationandcatalysis).
ThisworkhaspublishedinNatNanotechnol,2:577,2007andreceivedhighlypositivecommentsfrominternationalcolleagues:"Acatalyticpropertywidelyusedforlaboratorytestsandthetreatmentofwastewaterhasbeendiscoveredinironoxidenanoparticlesandcouldleadtomanyapplicationsinmedicaldiagnostics.
"(Hiddentalent,NatNanotechnol,2:535,2007).
发表文章:1、LengNie,LizengGao,XiyunYan,andTaihongWang.
Functionalizedtetrapod-likeZnOnanostructruresforDNAgenedelivery,SolidStatePhenomena121:747-750,20072、LizengGao,JieZhuang,LengNie,JinbinZhang,YuZhang,NingGu,TaihongWang,JingFeng,DonglingYang,SarahPerrettandXiyunYan.
Intrinsicperoxidase-likeactivityofferromagneticnanoparticlesandapplicationinanimmunoassay.
NatureNanotechnology.
2007;2(9):577-5833、PengchengBu,JieZhuang,JingFeng,DonglingYang,XunShenandXiyunYan.
VisualizationofCD146dimerizationanditsregulationinlivingcells.
BiochimicaetBiophysicaActa(BBA)-MolecularCellResearch.
2007;1773(4):513-5204、LinY,WuX,ShenY,BuP,YangD,YanX.
ANovelAntibodyAA98VH/LDirectedAgainstCD146EfficientlyInhibitsAngiogenesis.
AnticancerResearch.
2007;27(6):4219-2452(六)蛋白质药物与多肽药物1、梁伟组揭示了肿瘤发生发展的新机制:我们的研究发现,组织的纤维化为肿瘤细胞的生成和发展提供了有利的微环境,纤维化可促进正常细胞向肿瘤细胞转化及增值;纤维化为肿瘤细胞提供了适宜其定居和生长的微环境并促进肿瘤转移灶的形成.
TGF-β/Smad3信号通路在这一过程中起到关键作用可以作为药物筛选的新靶点.
发现了柚皮素和柚皮苷可以特异性干预TGF-β/Smad3信号通路起到预防和治疗纤维化及癌症作用.
提出了肿瘤化疗的新观点和新思路:大多数化疗药物的作用靶点在细胞内,如阿霉素和顺铂等作用于细胞核的DNA,紫杉醇和长春新碱等作用胞浆的微管;或已具有耐药性的肿瘤细胞,通过外排机制将药物泵出细胞外,细胞内的药物浓度不足以抑制细胞的增殖.
仅仅将药物输送到肿瘤组织(组织靶向)是不够的,在提高肿瘤组织选择性基础上,如何提高药物的组织渗透性及对肿瘤细胞选择性并能跨膜输送至细胞内达到有效浓度已成为肿瘤治疗的新的挑战.
我们提出了利用纳米药物输送系统既实现肿瘤组织的靶向渗透性又实现肿瘤细胞内的靶向富集,为临床治疗肿瘤提供新的有效手段和策略.
1、Prof.
LiangW.
RevealinganewmechanismoftumorgenesisandprogressionBasedonourresearch,wefindthattissuefibrosisprovidesaspecialmicroenvironmentwhichisbeneficialfortheproliferationoftumorcells.
Fibrosiscanpromotethemalignanttransformationofnormalcellsintotumorcells.
Fibrosisalsooffersabeneficialmicroenvironmentthatcanpromotetumorgrowthandmetastasis.
TGF-b/smad3signalingpathwayplaysacrucialroleintheseprocessesandcanbeanewtargetfordrugdesignanddevelopment.
NaringeninandNaringincanspecificallyinhibitTGF-b/smad3signalingpathwayandconsequentlycanbeusedforpreventionandtherapyoffibrosisandcancer.
ProposinganewviewpointtowardchemotherapyforcancerThetargetsofmostchemotherapeuticdrugwerelocatedintheintra-cells,suchasdoxorubicinandcisplatinintercalateintodoublehelixofDNAinthenucleus,andpaclitaxelandvincristindisruptthemicrotubesinthecytoplasma.
However,resistanttumorcellscanpumpdrugsoutofthecellsthroughsomespecificeffluxmechanisms.
Therefore,concentrationofthedrugincellscannotreachthecriticalleveltoinhibittumorcellproliferationorkilltumorcell.
Inordertoovercometheseproblems,itisnotenoughonlytodeliverdrugtothetumortissue(tissuetarget),butthedrugneedtobedeliveredintotumorcells.
Penetrationintothedepthoftumortissueandintotumorcellsreachingeffectiveconcentrationofdruginthetumorcellsisveryimportantforchemotherapy.
Wehavedevelopedanovelnano-sizeddrugdeliverysystemtofitfortherequirementsofpenetrationintothedepthoftumortissueandintotumorcells.
Thisstrategycouldhaveimportantclinicalapplicationsforcancertherapy.
发表文章:1、YiDD,LiGM,LiG,LiangW.
Interactionofarginineoligomerwithmodelmembrane.
BiochemicalandBiophysicalResearchCommunicatios.
2007;359(4):1024-10292、TangN,DuGJ,WangN,LiuCC,HangH,LiangW.
Improvingpenetrationintumorswithnano-assembliesofphospholipidsanddoxorubicin.
JournaloftheNationalCanerInstitute.
2007;99(13):1004-1015.
532、马跃组目前在人源化细胞外基质的研究中取得一定的进展,初步得到了一种可用于人胚胎干细胞培养的人源细胞外基质.
这对人胚胎干细胞的培养,移植具有一定的意义.
目前我们正在进一步的鉴定和优化.
2、Prof.
MaY.
WerecentlydevelopedahumanizedEMCforhumanembryonicstemcellculture.
ThisEMCareallhumanprotein,andcanavoidanimalproteincontainminationifappliedonhumanEScellculture.
WearecurrentlyworkingoncharacterizationofthisEMC.
543、殷勤伟组基础研究方面与中科院计算机所合作,研发了新的软件在曙光4000H生物信息处理的应用专用计算机上从2万1千多个人编码蛋白基因的内含子中发现了1千多个内源性shRNAs(shorthairpinRNAs),它们不同于现已发现的其它小RNAs如miRNAs,piRNAs,rasiRNAs,siRNAs等,是一类新的小RNAs,详细的研究成果正在整理、分析和发表过程中.
用这些新发现的小RNA和已发现的miRMNA研制成了小RNA组合芯片,已检测了6对正常的组织细胞和肿瘤细胞间的表达差异,发现它们也许是一种优良的标志物,可用于临床的诊断.
对此,已与其他研究组(如乐家昌研究员)合作进一步研制用于临床诊断的小RNA分子马达检测仪.
通过小RNA组合芯片,我们能够筛选出一些有重要功能的shRNAs,它们可通过多种方式来调控基因的表达和细胞的生长和分化.
对此,与中国协和医科大学合作,正在研究人内源性shRNA对人间充质干细胞的自我更新和生长分化的调控作用.
结果是令人兴奋的,第一阶段的研究成果正在整理、分析和发表过程中.
同时,我组也在小RNA与DNA甲基化方面作了一些研究工作,有关的研究论文正处于投稿阶段.
此外,我们也正在扩展和深入对这些新发现的小RNAs的功能和作用机制的研究,力争能做出一些原创性的重要发现3、Prof.
YinQ.
W.
BasicResearch1.
Withthecooperationwiththeinstituteofcomputingtechnology,wehavefoundmorethan1000novelsmallRNAsfromhumanintronsofprotein-codinggenes.
TheyaredifferentfromthoseknownsmallRNAssuchasmiRNA,tncRNA,rasiRNAs,piRNAsadothers.
2.
UsingthesenovelRNAsequences,wehavedevelopedanintegrativemicroarraysthatcontainknownmiRNAsandnewpredictedsRNAs.
Basedonthistypeofarrays,weassayed12differentsamplesincludingnormalandcancercells.
Furthermore,wearedevelopinganewmolecularengine-drivensmallRNAassayinstrumentwithotherlabs.
3.
Byusingsmallmicroarrays,mylabhasselectedsomeimportantsRNAsthatcanregulatethedifferentiationandgrowthofcells.
Forexample,ofthemtwosRNAscanmodulatetheproliferationanddifferentiationofhaematopoieticstemcells.
4.
WealsoconductedsomeinvestigationsinsRNAsandDNAmethylation.
Theresearchdataarebeingpreparedforpublication.
5.
Moreimportantly,mylabisgoingtomakedeeperandbroaderstudyonthebiogenesisandcellularfunctionsofthesenewlydiscoveredsRNAs.
发表文章:TangY,GeYZ,YinQW.
ExploringinvitrorolesofsiRNAincardiovasculardisease.
ActaPharmacolSin.
200728(1)55六、研究成果(Achievements)1、2007年度发表论文列表(2007Publications)序号论文课题组1DK,KWANGZHAOJ,ASOGYONG,YANGXM,DUPS,TANGH,FUYX,Adaptiveimmunecellstemperinitialinnateresponses.
NATUREMEDICINE.
2007,1248-1252唐宏2ZHIJUNZHANG,GANGCHEN1,WEIZHOU1,AIHONGSONG,TAOXU,QINGMINGLUO,WEIWANG,XIAO-SONGGUANDSHUMINDuan1RegulatedATPreleasefromastrocytesthroughlysosomeexocytosis,NAURECELLBIOLOGY.
2007,449:945-953徐涛3BAIL,WANGY,FANJM,CHENY,JIW,QUAL,XUPY,JAMESDE,XUT,DissectingmultiplestepsofGLUT4traffickingandidentifyingthesitesofinsulinaction.
CELLMETAB.
2007,5(1)47-57.
徐涛4TANGN,DUGJ,WANGN,LIUCC,HANGHY,LIANGW,Improvingpenetrationintumorswithnanoassembliesofphospholipidsanddoxorubicin.
JNATCANCERINST.
2007,99(4)1004-1015梁伟5KE-MINGZHOU;YONG-MINGDONG;QIANGE;DANZHU;WEIZHOU;XIAN-GUANGLIN;TAOLIANG;ZHENG-XINGWU;TAOXU,PKAactivationbypassestherequirementforUNC-31inthedockingofdensecorevesiclesfromC.
elegansneurons.
NEURON.
2007,56:657-669徐涛6YONGQUNZHU,HONGTAOLI,CHENGZULONG,LIYANHU,HAOXU,LIPINGLIU,SHECHEN,DA-CHENGWANG,FENGSHAO,StructuralInsightsintotheEnzymaticMechanismofthePathogenicMAPKPhosphothreonineLyase,MOLECULARCELL.
2007,28:899-913王大成7NEILSHAW,MINZHAO,CHONGYUNCHENG,HAOXU,JUHASAARIKETTU,YANGLI,YURONGDA,ZHIYAO,OLLISILVENNOINEN,JIEYANG,ZHI-JIELIU,BI-CHENGWANG&ZIHERAO,Themultifunctionalhumanp100protein'hooks'methylatedligands.
NATURESTRUCTURAL&MOLECULARBIOLOGY.
2007,14:779-784刘志杰8GuangyuZhu,JiaChen,JayLiu,JosephSBrunzelle,BoHuang,NancyWakeham,SimonTerzyan,XuemeiLi,ZiheRao,GuangpuLiandXuejunCZhang,StructureoftheAPPL1BAR-PHdomainandcharacterizationofitsinteractionwithRab5.
EMBOJ.
2007,26:3484–3493饶子和9LPAN,SYCHEN,CJWENG,GCALL,DXZHU,HTANGANDTXIEStemcellagingiscontrolledbothintrinsicallyandextrinsicallyinthedrosophilaovary,CELLSTEMCELL.
2007,1:458-469唐宏10GAOLZ,ZHUANGJ,NIEL,ZHANGJB,ZHANGY,GUN,WANGTH,FENGJ,YANGDL,PERRETTS,YANXY,Intrinsicperoxidase-likeactivityofferromagneticnanoparticles.
NATURENANO.
2007,2:577-583阎锡蕴11GUOX,MAJ,SUNJ,GAOGXThezinc-fingerantiviralproteinrecruitstheRNAprocessingexosometodegradethetargetmRNA.
PANS2007,104(1)151-156.
高光侠12JINZHONGLIN,,TAOZHOU,KEQIONGYE,,ANDJINFENGWANG,Crystal王金凤56structureofhumanmitoNEETrevealsdistinctgroupsofiron–sulfurproteins.
PROCNATLACADSCIUSA.
2007,104:14640-1464513ZHAO,X.
,M.
MOHAUPT,J.
JIANG,S.
LIU,B.
LI,ANDZ.
QIN.
TumorNecrosisFactorReceptor-2-MediatedTumorSuppressionisNitricOxideDependentandInvolvesAngiostasis.
CANCERRE.
2007,67(9):4443-50.
秦志海14SHE,CNLIU,GSKOGERB,HTZHAO,JWANG,TLIU,BYBAI,YZHAOANDRSCHEN,NONCODEv2.
0:decodingthenon-coding,NUCLEICACIDSRESEARC.
,2007,1–3陈润生15ZHAOT,ZHANGH,GUOY,ZHANGQ,HUAG,LUH,HOUQ,LIUH,FANZGranzymeKcleavesthenucleosomeassemblyproteinSETtoinducesingle-strandedDNAnicksoftargetcells.
CELLDEATHDIFFER2007,14(3)489-499范祖森16BUPC,ZHUANGJ,FENGJ,YANGDL,SHENX,YANXY,VisualizationofCD146dimerizationanditsregulationinlivingcells.
BBA-MOLCELLRES2007,513-520阎锡蕴17JIANJUNCHEN,LUCHEN,GANGWANG,HONGTANGCholesterol-Dependentand-IndependentCD40InternalizationandSignalingActivationinCardiovascularEndothelialCells.
ARTERIOSCLERTHROMBVASCBIOL.
2007,12:17626904唐宏18JIAOR,HARRIGANJA,SHEVELEVI,DIETSCHYT,SELAKN,INDIGFE,PIOTROWSKIJ,JANSCAKP,BOHRVA,STAGLJARI,TheWernersyndromeproteinisrequiredforrecruitmentchromatinassemblyfactor1followingDNAdamage.
ONCOGENE.
2007.
26:3811-3822.
焦仁杰19QU,JING;LIU,GUANG-HUI;HUANG,BO;CHEN,CHANG,NitricoxidecontrolsnuclearexportofAPE1/Ref-1throughS-nitrosationofCysteines93and310.
NUCLEICACIDSRESEARCH.
2007,35:2522-2532陈畅20LI,Z.
,P.
FELICIA,B.
LI,S.
LIU,ANDZ.
QIN.
CrosstalkbetweenTcellsandinnateimmunecellsiscrucialforIFNγ-dependenttumorrejection.
JIMMUNOL.
2007,179(3):1568-76.
秦志海21CHENLG,ZHANGZ,CHENWW,ZHANGZD,LIYG,SHIM,ZHANGJY,CHENLP,WANGSD,WANGFS,B7-H1up-regulationonmyeloiddendriticcellssignificantlysuppressesTcellimmunefunctioninpatientswithchronichepatitisB.
JIMMUNOL2007,178(10):6634-41王盛典22WANGR,YINYJ,WANGF,LIM,FENGJ,ZHANGHM,ZHANGJP,LIUSJ,ChangWR.
Crystalstructuresandsite-directedmutagenesisofamycothiol-dependentenzymerevealanovelfoldingandmolecularbasisformycothiol-mediatedmaleylpyruvateisomerization.
JBIOLCHEM.
2007,282(22):16288-294.
常文瑞23JBENACH,LWANG,YCHEN,CKHO,SLEE,JSEETHARAMAN,RXIAO,TBACTON,GTMONTELIONE,HDENG,RSUN,ANDLTONG.
StructuralandfunctionalstudiesoftheabundanttegumentproteinORF52frommurinegammaherpesvirus-68.
JOURNALOFBIOLOGICALCHEMISTRY2007,282:31534-31541,邓红雨24HUAGQ,ZHANGQX,FANZS,Heatshockprotein75(TRAP1)antagonizesreactiveoxygenspeciesgenerationandprotectscellsfromgranzymeM-mediatedapoptosis.
JBIOLCHEM2007,282(28)20533-20560范祖森25TONGBIAOZHAO,HONGLIANZHANG,YUMINGGUO,ANDZUSENFAN.
GranzymeKdirectlyprocessesbidtoreleasecytochromecandendonucleaseG范祖森57leadingtomitochondria-dependentcelldeath.
JBIOLCHEM.
2007,282(16):12104-1126HOUX,LIUR,ROSSS,SMARTEJ,ZHUH,GONGW.
CrystallographicStudiesofHumanMitoNEET.
JBIOLCHEM.
2007,282(46):33242-6.
龚为民27IMMELF,JIANGY,WANGYQ,MARCHALC,MAILLETL,PERRETTS&CULLINCInvitroanalysisofSpUre2p,aprionrelatedprotein,exemplifiestherelationshipbetweenamyloidandprion.
J.
BIOL.
CHEM.
2007,282:7912-7920.
柯莎28LIAN,H.
Y.
,ZHANG,H.
,ZHANG,Z.
R.
,LOOVERS,H.
M.
,JONES,G.
W.
,ROWLING,P.
,ITZHAKI,L.
S.
,ZHOU,J.
M.
&PERRETT,S.
Hsp40interactsdirectlywiththenativestateoftheyeastprionproteinUre2andinhibitsformationofamyloid-likefibrils.
J.
BIOL.
CHEM.
2007,282:11931-11940.
柯莎29YES,WUX,WEIL,TANGD,SUNP,BARTLAMM&RAOZ.
Aninsightintothemechanismofhumancysteinedioxygenase:Keyrolesofthethioether-bondedtyrosine-cysteinecofactor.
JBIOLCHEM.
,2007,282(5):3391-3402饶子和30ZHAOQ,QINL,JIANGF,WUB,YUEW,XUF,RONGZ,YUANH,XIEX,GAOY,BAIC,BARTLAMM,PEIX*&RAOZ.
Structureofhumanspindlin1:tandemtudor-likedomainsforcellcycleregulation.
J.
BIOL.
CHEM.
,2007,282(1):647-656饶子和31WANGZX,WUJW,ThecompletepathwayforERK2-catalyzedreaction-EvidenceforanisorandomBiBimechanism.
JBIOLCHEM.
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X用于检测葡萄糖转运蛋白4上膜的探针和方法徐涛11200710100124.
9检测过氧化氢含量的新试剂和新方法阎锡蕴12EP2007/008961TheproteinfactorLepA(EF4)asanewtargetforantibioticsagainstbacteria秦燕3、2007新增国家、院级重要主持项目(2007Projects)序号项目来源项目编号项目/课题名称主持人1创新研究群体30721003膜蛋白与蛋白质复合体的结构生物学研究常文瑞2海外青年学者合作研究30728004生物化学和分子生物学朱海宁龚为民3海外青年学者合作研究基金30728006免疫学程根宏唐宏4国际合作重点2007DFC30190表型组学技术:病毒感染免疫应答机制中的应用唐宏5863重大项目2006AA02A245肿瘤抗体药物阎锡蕴6863重大项目2006AA02A106心血管疾病干细胞临床治疗技术与产品的研发马跃7863重大项目2006AA02A314癌症相关蛋白质的三维结构研究刘迎芳8863重大项目2006AA02A319心血管、神经与免疫系统重大疾病相关蛋白的三维结构研究江涛9院重大项目KSCX1-YW-02-12型糖尿病致病机制及新药靶徐涛10院重大项目KSCX1-YW-10艾滋病和病毒性肝炎新型疫苗和新药研究唐宏11院重大项目KSCX1-YW-13冷冻电镜三维重构原始数据集孙飞12院方向性项目KSCX2-YW-R-120逆转录病毒与宿主相互作用机理研究高光侠13院方向性项目KSCX2-YW-R-121肿瘤抗体药物阎锡蕴6314院方向性项目KSCX2-YW-R-123物质运送、能量转换相关膜蛋白的结构与功能研究常文瑞15院方向性项目KSCX2-YW-R-125HOPS蛋白复合体在线虫细胞凋亡中作用的分子机制研究刘迎芳16院方向性项目KSCX2-YW-R-126冷冻电子断层三维成像技术建立及其应用孙飞17院方向性项目KSCX2-YW-R-127动化高效率晶体结构解析系统的研究和构建刘志杰18院仪器研制改造项目Y20634高通量、高灵敏度蛋白质组学研究系统杨福全19院仪器研制改造项目YZ200751光纤型ATP马达生物传感器miRNA分析仪器的研制殷勤伟64七、研究生培养(Training)(一)博士后2007年在站人博士后:24人2007年出站人博士后:3人名单:闫小雪、徐平勇、张志栋(二)博士生2007年在读博士生:139人2007年取得博士学位:27人名单:马静、张佰茹、耿勇、白宝艳、卜鹏程、曹鹏、曹晟、陈桂芳、韩佩韦、韩伟、郝蕊、侯强、黄文涛、霍霞、李梅、连惠勇、刘涛、吕红霞、欧先金、曲静、苏华、孙红、孙阳、宛佳、王铁鹏、殷雷、张红、朱小蓬、朱永群(三)硕士生2007年在读硕士生:162人2007年取得硕士学位:4人名单:张玲、李琳、李绍娟、李文奇(四)学生获奖卜鹏程、刘光慧、孙红院长优秀奖曲静刘永龄奖何云、谢韬地奥奖学金一等奖牛玉琼、王峰地奥奖学金二等奖赵学强研究生院BHPBilliton学生奖学金庄洁第九届复旦大学谈家桢基金九源奖学金一等奖王琰、赵洁、贺子轩、林金钟、孙红第九届复旦大学谈家桢基金九源奖学金三等奖八、学术交流(InternationalExchange)(一)承办的学术会议1、中国科学院结构生物学战略研讨会时间:2007-4-5地点:北京2、基础与应用免疫生物学国际研讨会时间:2007-10-1地点:天津(二)学术交流参与国内外学术会议:76人次,会议报告:41人次国内外来室讲学:56人次国内外来室访问:42人次65九、2007年度大事记(2007Events)一月1.
17古巴科学院院士、中巴双边生物技术工作组组长、古巴神经科学中心主任MitchellValdes-Sosa博士在古巴驻中国大使馆一等秘书HectorCondeAlmeida先生的陪同下到生物物理研究所进行学术访问并参观了实验室.
1.
21第六届中国十大女杰评选活动在京揭晓,中国科学院院士、发展中国家科学院院士、生物大分子国家重点实验室学术委员会主任中国科学院生物物理研究所研究员王志珍同志光荣当选.

二月2.
15中国科学院副院长陈竺看望了全国政协委员、九三学社中央副主席、中国科学院院士、第六届中国十大女杰获得者、生物大分子国家重点实验室学术委员会主任王志珍院士.

三月3.
20在2007年1月召开的北京市科学技术协会第七次代表大会上,生物大分子国家重点实验室学术委员会主任王志珍院士当选为北京市科协第七届委员会委员、副主席.

3.
20中国科学院青年联合会二届三次常委会议在北京召开,会议增补了生物大分子国家重点实验室主任徐涛研究员为第二届中科院青联副主席.
3.
22德国科学基金会(DFG)化学学部主任Dr.
KarlheinzSchmidt在德国科学基金会赵妙根主任的陪同下访问生物物理研究所并参观了实验室.
3.
22科技部国际合作司马林英副司长在中科院国际合作局曹京华副局长、科技部国际合作司综合与计划处张健处长、美大处王强处长的陪同下视察生物物理研究所参观唐宏研究员的课题组.
生物物理研究所党委书记杨星科、副书记实验室副主任龚为民等参加了接待.

四月4.
10来自17个国家的30余位驻华使馆科技官员和国际组织驻华代表到生物物理研究所和实验室进行参观访问.
4.
14由生物物理研究所和生物大分子国家重点实验室共同承办的"中国科学院结构生物学战略研讨会"在香山召开.
陈竺院士、饶子和院士、徐涛研究员担任本次会议执行主席,来自海内外的30多位知名结构生物学家以及物理学、化学等领域的专家参加了本次会议,科技部、基金委等部门的领导应邀出席了会议.
4.
19蛋白质科学国家实验室筹备办公室正式挂牌.
五月5.
19第七届全国科技活动周和中科院第三届公众科学日活动开幕式在北京奥运村中科院动物所开幕.
当日,由生物物理研究所主办的以"关注生命科学,共建和谐社会"为主题的第四届"公众科学日"同时拉开帷幕.
实验室承接了来自科研院所、大学、中小学及社会各界群众200余人参加.
66六月6.
11我室徐涛研究员承担中科院仪器改造项目"膜蛋白实时相互作用的荧光共振能量转移动态检测系统"顺利通过院综合计划局专家组验收.
6.
21德国马普学会科学家代表团到生物物理研究所和生物大分子国家重点实验室进行学术访问.