肝脏蛋白质组计划的实施与毒理学发展的机遇演示文稿

肝脏蛋白质组计划的实施与毒理学发展的机遇演示文稿目前一页\总数一百一十三页\编于十五点（优选）肝脏蛋白质组计划的实施与毒理学发展的机遇目前二页\总数一百一十三页\编于十五点曼哈顿原子弹研制计划人类基因组计划阿波罗登月计划人类历史上的三大科技工程罗斯福批准,耗资20亿美元原子半径10-10m原子体积10-30m3人体半径100m人体体积100m3太阳系半径1012m太阳系体积1034m3克林顿、布莱尔批准，耗资30亿美元肯尼迪批准,耗资240亿美元目前三页\总数一百一十三页\编于十五点人类基因组计划开创了

“基因组时代”目前四页\总数一百一十三页\编于十五点基因组学向蛋白质组学“求助”!

Nature409:747,15Feb.2001

Andnowforproteome

“现在轮到蛋白质组”Science291:1221,16Feb.2001ProteomicsinGenomeland“基因组大地中的蛋白质组学”目前五页\总数一百一十三页\编于十五点PROTEIN目前六页\总数一百一十三页\编于十五点转录组一种细胞、组织或生物体所对应的全套mRNARNA基因组生物体所拥有的全套染色体上的全部基因DNA蛋白质组一种细胞、组织或生物体所对应的全套蛋白质PROTEIN功能执行体遗传信息载体目前七页\总数一百一十三页\编于十五点

基因和蛋白质并不存在严格的线性关系

ORF并不预示一定存在相对应的功能性基因

mRNA水平并非与蛋白质的表达水平对应翻译后修饰及同工蛋白质(Isoforms)等现象在基因水平无从表现蛋白质与蛋白质的相互作用难以在基因水平得以认识基因组与转录组不能取代蛋白质组目前八页\总数一百一十三页\编于十五点翻译后修饰相互作用构象变化翻译后拼接移位胞质胞核蛋白质调节的多样性生命的“万花筒”目前九页\总数一百一十三页\编于十五点蛋白质功能的群体性目前十页\总数一百一十三页\编于十五点绳墙扇茅蛇树蛋白质作用的整体性目前十一页\总数一百一十三页\编于十五点探索生命奥秘的直接对象蛋白质组

生命体的统一性源于基因组生命体的多样性、复杂性、功能性、表型源于蛋白质组1463-4x1043x109103(1012)

目前十二页\总数一百一十三页\编于十五点

Pteomicsismoreofaconceptthanadefinedtechnology,anditreferstoanyprotein-basedapproachthathasthecapacitytoprovidenewinformationaboutproteins

onagenomewidescale.“Proteomicsincludesnotonlytheidentificationand

quantificationofproteinsbutalsothedeterminationof

their:

localization

modifications

interactions

activities

function目前十三页\总数一百一十三页\编于十五点报告提要蛋白质组学产生的时代背景“国际人类肝脏蛋白质组计划”的目标与意义蛋白质组学与毒理学/环境医学目前十四页\总数一百一十三页\编于十五点DNA序列图≠基因图人类基因组中1/3以上基因未曾确认基因确认的基本层次--蛋白质水平天书解读天书Science291:1221,2001目前十五页\总数一百一十三页\编于十五点全面揭示重大疾病发生发展机制的基础蛋白质组单基因病—遗传病多基因病—肿瘤等

重大疾病发生发展机制单一基因单一蛋白基因群蛋白质群转录组蛋白质组????目前十六页\总数一百一十三页\编于十五点发现大量新型药靶与新药的源泉蛋白质组最重要的疾病100-150每种疾病相关的基因10每个基因相关的蛋白质3–10药靶蛋白总数

3000-15,000

现有药物约2000种85％是针对目前已知的500种药靶6-30倍药靶有待发现目前十七页\总数一百一十三页\编于十五点启动人类蛋白质组计划势在必行！探索人体生命奥秘的直接对象全面揭示重大疾病发生发展机制的基础发现大量新型药靶与新药的源泉

人类基因组序列及基因注释人类蛋白质组VIP:VeryImportantProteome目前十八页\总数一百一十三页\编于十五点人类蛋白质组计划的

主要科学目标验证人类基因组计划推测的基因，注释基因组阐明蛋白质组的调控模式并与转录组进行对比建立蛋白质群/组装体，蛋白质复合物或蛋白质机器,即连锁图建立人体生理学、病理学的蛋白质组基础目前十九页\总数一百一十三页\编于十五点1463-4x1043x109103(1012)

基因组计划蛋白质组计划目前二十页\总数一百一十三页\编于十五点目前二十一页\总数一百一十三页\编于十五点Bodyfluidmostaccessibleforbiomarkerdiscoveryinanimalandhumans.LiverKidneyOrganBodyFluidBrainImmuneOrgans/VesselsIntestineOtherTissuesCSFFecesEndocrine/ParacrineSecretionsBileUrineAir/BALFLymphLungsEyesSkinOralCavitySweatSalivaTearsVenousBloodArterialBloodSerum10%90%SerumProteinsNon-InformativeAbundantProteinsi.e.albumin,IgG,etc.InformativeLowAbundanceProteinsEnrichmentStrategiesforInformativeSerumProteins:SELDI

SerumImmunosubtraction

目前二十二页\总数一百一十三页\编于十五点人类肝脏蛋白质组计划里程碑蛋白表达谱蛋白连锁图亚细胞定位图修饰谱人类基因组计划

里程碑遗传图物理图序列图目前二十三页\总数一百一十三页\编于十五点InitiativesofHumanProteomeProject(HPP)HPPPHumanPlasmaProteomeProject,USAHLPPHumanLiverProteomeProject,ChinaPSIProteomicsStandardsInitiativeUKHBPP

HumanBrainProteomeProject,GermanyHAIHumanAntibodyInitiativeSweden目前二十四页\总数一百一十三页\编于十五点Whyisliver?wwwww.HLPP.HUPO为何研究肝脏?目前二十五页\总数一百一十三页\编于十五点肝脏功能的多样性与重要性能量——能量转换、储存物质——代谢（活性分子的合成、毒性分子的分解）信息——信息分子的合成与分泌目前二十六页\总数一百一十三页\编于十五点肝脏为其它组织提供能量—可氧化底物组织的活力依赖于高能键的连续生成。肝脏产生大部分脂肪酸，后者是禁食状态下能量的基本来源。肝脏是给食状态下由过剩糖合成脂肪酸的主要部位。目前二十七页\总数一百一十三页\编于十五点药物毒物营养物生物异源物生物转化目前二十八页\总数一百一十三页\编于十五点化学多样性向体内的生物转化体系提出严峻挑战1.2×107种以上的化学物(CAService’slist)以每周约8000种的速率增加常用化学物在63,000种以上大约11,500种，作为食物或药物制剂添加物摄入其余的50,000种，为潜在的环境污染物目前二十九页\总数一百一十三页\编于十五点人类发育过程中造血组织的兴替造血系统的发育必需肝脏的“培育”目前三十页\总数一百一十三页\编于十五点合成大多数循环血浆蛋白合成和分泌血浆蛋白是肝脏实质细胞——肝细胞的独有功能（肝细胞占肝脏总细胞数的60%及肝脏质量的约80%）最有特征的血浆蛋白是白蛋白，在大多数哺乳动物中占总血浆蛋白的55-60%凝血酶、抗凝因子、溶栓酶等目前三十一页\总数一百一十三页\编于十五点肝脏再生机体再生能力最强的器官终生保持旺盛的再生能力目前三十二页\总数一百一十三页\编于十五点肝脏中包含的“组”(-ome)Metabolicgenomics→Metabolome

(代谢组)Energygenomics→Energome

(能量组)Pharmacogenomics→Pharmacome

(药理组)Toxicogenomics→Toxicome

(毒理组)Regeneratinggenomics

→Regenerome

(再生组)目前三十三页\总数一百一十三页\编于十五点全球及中国肝炎的流行病学统计

全球:3.5亿携带者中国:1.8亿携带者死亡:23万人/年疾病负担:500亿元人民币治疗手段:抗病毒,保肝降 酶,抗纤维化,免疫治疗等缺点:病毒易反弹,病 情易反复目前三十四页\总数一百一十三页\编于十五点肝炎向肝癌的恶性转化

难以遏制全世界现有3.5亿慢性乙肝病毒（HBV）携带者，占世界人口的5%，亚洲和非洲HBV携带率为8-15%

HBV携带者中50-70%病毒复制活跃，为慢性乙肝

慢性乙肝病人肝硬化发生率为2-20%，代偿性肝硬化发展成失代偿性肝硬化为20-23%，发展成肝癌的占6-15%中国的慢性乙肝病人中，约25-40%最终将死于肝硬化或合并肝癌HBV携带者最终死于相关肝病的危险性，男性为50%，女性为15%

目前三十五页\总数一百一十三页\编于十五点全球:100万人/年中国:50万人/年死亡:约45万人/年疾病负担:400亿元人民币治疗手段:手术切除(只有 15%-20%)治疗效果:术后易转移,易复 发,五年存活率 40-50%全球及中国肝癌的流行病学统计目前三十六页\总数一百一十三页\编于十五点原发性肝癌的治疗水平亟待突破世界上每年有100万新发原发性肝癌病人，其中40-50%在我国

我国原发性肝癌发病率和死亡率均位居第二位

近20年来我国肝癌的发病率上升近40%原发性肝癌一般起病较隐匿，早期缺乏典型临床表现，初诊大多已属中晚期

初诊肝癌病人中，只有15-20%的病人具备手术条件这些病人术后5年生存率仅为40-50%

肝癌是致死率最高的恶性肿瘤之一目前三十七页\总数一百一十三页\编于十五点重大肝病的发生发展无法归咎于少数几个基因或蛋白质所呈现的功能状态和信号通路从蛋白质组层面上“全景式”揭示肝脏疾病的生理病理机制是解决重大肝病的根本出路肝炎病毒肝脏/肝细胞肝炎肝硬化/纤维化原发肝癌肝癌转移多因素、多步骤的发病机制目前三十八页\总数一百一十三页\编于十五点肝脏是人类蛋白质组计划的首批目标！目前三十九页\总数一百一十三页\编于十五点Prometheusstolefirefromthegodsandgaveittomortals.PrometheusBoundHeraclesLiberatePrometheusHLPP—ModernPrometheusMythPrometheus

LiverEagleLiverdiseases

HeraclesHLPP目前四十页\总数一百一十三页\编于十五点人类肝脏蛋白质组计划

HumanLiverProteomeProject(HLPP)国际HLPP共同体目前四十一页\总数一百一十三页\编于十五点Generateanintegrativeapproachleadingtoacomprehensivefunctionalmapoftheliver

Expandliverproteometoits“PHYSIOME”and“PATHOME”todramaticallyacceleratethedevelopmentofdiagnostics,preventionandtherapeuticstowardsitsdiseases

Developstandardoperatingprocedures(SOPs)forotherHUPOInitiativesVISION目前四十二页\总数一百一十三页\编于十五点HUPOWorkshop,Bethesda,April28-29,2002

BroadinterestsandsupportsforinitiatingHLPP

HUPOLiverProteomeWorkshop,Beijing,October22-24,2002

GettingconsensusviewforscientificobjectivesofHLPP

HUPO1stWorldCongress,Versailles,November21-24,2002

Co-chairsofHLPP:Drs.FuchuHe,JohnBergeronandChristianBréchot

HLPPPlanningCommittee:17members;May2003ProposalsaboutWorkingPlanofHLPP

Dr.JohnBergeron:Jan31,2003(Management);

Dr.FuchuHe:Feb17,2003(ScientificStrategy)Dr.FuchuHe:Mar22,2003(ActionPlan);

Dr.ChristianBrechot:Mar31,2003(Sampling)

HLPPOffice,March18,2003HUPOLPPWorkshop,Bethesda,July17-18,2003

NIHparticipatingHUPO2ndWorldCongress,Montreal,Oct.8-12,2003

Dr.FuchuHewasappointedastheexecutiveChairofHLPP(2003-2005)HUPOHLPPSatelliteMeeting,Montreal,Oct.12,2003SetupExecutiveCommittee&9Sub-committees,ActionPlanatPilotPhaseFrenchliversampleswerecollectedanddistributedamong8labs,May,2004HUPOHLPPSatelliteMeeting,Beijing,Oct.24,2004ChinesepartofHLPPwasofficiallylaunched,Nov.,2004Chineseliversampleswerecollectedanddistributedamong7labs,Feb.,2005MISION目前四十三页\总数一百一十三页\编于十五点表达谱修饰谱定位图连锁图样品库抗体库数据库HLPP的科学目标---肝脏蛋白质组的“太阳系”3-D图ORF组目前四十四页\总数一百一十三页\编于十五点肝脏蛋白质组与

肝脏生理、病理的衔接肝脏蛋白质组代谢组毒理组药理组再生组肝炎组肝癌组能量组目前四十五页\总数一百一十三页\编于十五点作为分泌器官，肝脏合成大多数的循环血浆蛋白

肝脏转录组“肝脏蛋白质组计划”和“血浆蛋白质组计划”的整合肝脏蛋白质组血浆蛋白质组?目前四十六页\总数一百一十三页\编于十五点基因组肝脏蛋白质组肝脏转录组基因组、转录组和蛋白质组的集成证实推测基因对比调控模式

目前四十七页\总数一百一十三页\编于十五点重要功能蛋白质肝病药物靶标肝病诊断标志物人类肝脏蛋白质组目前四十八页\总数一百一十三页\编于十五点肝炎病毒肝脏/肝细胞感染肝炎肝硬化/纤维化原发肝癌肝癌转移重大肝病预警、诊断、预防、治疗的关键环节“东亚病夫”“肝病大国”1亿多肝炎病毒携带者1000亿多/年医疗费用目前四十九页\总数一百一十三页\编于十五点SOPsSpecimenBankingPlatformsWorkingteamGlobalCollaborationExpressionProfileORFeomeAntibodiesBioinformatics…

ModificationProfileSubcellularLocalizationProtein-ProteinInteractionAntibodiesBioinformaticsIntegrateproteomewithtranscriptomeCompareproteomeswithinhealthanddiseasedliverTimeLinesPilotPhase(2003-2005)PhaseII(2006-2010)目前五十页\总数一百一十三页\编于十五点OutlineBackgroundProgressoftheHLPP-ProgressreportonexpressionprofilingInitiationoftheCNHLPPNextwork目前五十一页\总数一百一十三页\编于十五点Progress-ExpressionprofilingEvaluationoftechnologiesBioinformaticsset-upSamplingandpreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues目前五十二页\总数一百一十三页\编于十五点ProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues目前五十三页\总数一百一十三页\编于十五点Majorconclusionsfromthevaluationoftechnologies–16standardproteins/7labsHighmolecularweightandlowabundanceproteinsarelessdetectableby2DE.Proteinswithonlythreeorderofmagnitudecouldbeidentifiedin2DE,butproteinswithfourorderofmagnitudecouldbeidentifiedinshotguntechnology.Morepeptides(redundant)wereidentifiedforhighMWproteinsinshotguntechnology.Somesimilarproteinswereidentified(notproteingroup),suggestingthecriteriafordistinguishingsimilarproteinsshouldbemorestrict.目前五十四页\总数一百一十三页\编于十五点ProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues目前五十五页\总数一百一十三页\编于十五点HLPPExperimentalDatabaseMWSDataClient…GUI，WebServices,APIFilesInterchange,DMBSDumpDatasynchronizeDatasubmissionExperimentreports,data,files…

HLPPDataMirrorMWSDataClientMWSDataClientMWSDataClientMWSDataClientMWSServerAdministratorHLPPDataMirrorSubmissionPackagesRepositoryHLPPProjectManagementDatabaseHLPPProjectManagementSystemHLPPParticipatingLaboratoriesHLPPDataCenterTheHLPPDataManagementSystem

ArchitectureBasedonMyWorkSpaceSystem(MWS)目前五十六页\总数一百一十三页\编于十五点CurrentStandarddraftofHLPPSampleInformationProteinextractionConcentrationMeasurementSeparationGel-based2DEGel1D-IEFGel1D-SDSLC-basedRPLCSCXLCSAXLCSECTrapDesaltingDigestionIonsourceMALDIAutoflexUltraflexABI4700ESIQstarQTofMicroLCQLTQMassspectrometryTOFTOFTOFUltraflexABI4700QTOFQstarQTofMicroITMSLCQLTQPeaklistgenerationABI4700QStarAutoflexTurboSequestMasslynxFlexAnalysisPeaklistpreprocessingGPSpeaklistpreprocessProtein/peptideidentificationMascotSequestProteinlistgenerationAutoflexGPSBiotoolsQstarBuildSummaryDTASelectBioworksAutoQuest目前五十七页\总数一百一十三页\编于十五点Proteinidentificationisbasedontheanalysisofpeptidesgeneratedbyproteolyicdigestion.Whenthedetectedpeptidesmatchmanyproteinsandcannotidentifyauniqueprotein,theresultwillbepresentedintheformofagroupofpossibleproteins.

DefinitionofProteinGROUP

目前五十八页\总数一百一十三页\编于十五点CriteriaforproteinidentificationIontrapXCorr(highest)≥1.9(1),2.2(2),3.75(3);DeltaCn≥0.1;RSp≤4.ABI-TOF/TOFProteinscore≥59(Rank1forthespot)Q-TOForQ-STARProteinscore>thresholdPeptidescore>“thescoreforidentity”or29MALDI-TOFProteinscore>thresholdTheproteinsinfirstreport(multi-proteinforthemixture)目前五十九页\总数一百一十三页\编于十五点ProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandpreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues目前六十页\总数一百一十三页\编于十五点EvaluationoftheSOPsforthesubcellularfractionationObject:FindtheoptimummethodofpretreatmentSample:LivertissuesofC57miceSubcellular:Mitochondria,Golgi,PM,Nucleus,ER,CytoplasmSamplepretreatmentFreshtissuesFrozenhomogenisedtissuesFrozentissues目前六十一页\总数一百一十三页\编于十五点Purity(Westernblotting):Freshtissues>frozenhomogenizedtissues>frozentissuesTheyieldofproteins:Freshtissues>frozenhomogenizedtissues>frozentissuesIntegrity(TEM):Freshtissues>frozenhomogenizedtissuesSummary目前六十二页\总数一百一十三页\编于十五点ProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues目前六十三页\总数一百一十三页\编于十五点ItisagiantambitiousobjectiveandgivesagreatchallengetoembryonicproteomicsItisnecessarytocarryoutapreviewbeforetheformallaunchingofthisprojectScience302:1316,Nov.21,2003Nature425:441,Oct.2,2003目前六十四页\总数一百一十三页\编于十五点AverageThroughputofthePlatform2DESystem

12Gel/3DaysAuto-SpotDigestSystem

1152spots/DayMSandMS/MSSystem

500-1000Spots/Day目前六十五页\总数一百一十三页\编于十五点TheflowchartofCCPITfortheproteinexpressionprofileofhumanfetalliver目前六十六页\总数一百一十三页\编于十五点Theareascalesbeforeandafterisoformprocessingingroupandproteinlevelsrespectively.Inproteinlevel,theredcircularityareasrepresentthenumberofconfirmedproteinsandtheyellowannulusareasrepresentthenumberofpossibleproteins.Non-isogroups19%Extensivelyconfirmed81%Groups41%Confirmed59%Non-isoforms43%Isoforms57%contribute

545

additionalproteins

and

eliminate763

groupscontribute

545

additionalproteins

and

eliminate1335

possible

proteinsIn

Protein

LevelIn

Group

Levelconfirmed:1950possible:

2593extensivelyconfirmed:2495non-isoformpossible:

1258目前六十七页\总数一百一十三页\编于十五点ACBChromosomaldistributionofHFLproteome-encodinggenes目前六十八页\总数一百一十三页\编于十五点FunctionalModulesinplasmamembrane目前六十九页\总数一百一十三页\编于十五点ProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues目前七十页\总数一百一十三页\编于十五点

ReferenceLabsforHLPPExpressionProfilingProject

FuchuHe&XiaohongQian(BeijingInstituteofRadiationMedicine)RongZeng(ShanghaiInstituteforBiologicalSciences)PengyuanYang(FudanUniversity)SiqiLiu(BeijingGenomicsInstitute,ChineseAcademyofSciences)目前七十一页\总数一百一十三页\编于十五点LauraBeretta(FHCRC,USA)RichardJ.Simpson(RoyalMelbourneHospital,Australia)

AngelikaGorg(TechnischeUniversitatMunchen,Germany)MarkBaker/JunhongSun(APAF,Australia)目前七十二页\总数一百一十三页\编于十五点OutlineBackgroundProgressoftheHLPPInitiationoftheCNHLPPNextwork目前七十三页\总数一百一十三页\编于十五点CNHLPPOverviewThefirstphase:2004-2005Fundingfromcentralgovernment:10millionUSdollars ChineseMOST:jointlyfundedbyNationalProgramon KeyBasicResearchProjects(973),NationalHigh-tech R&DProgram(863),andNationalKeyTechnologiesR&D Programmeininthefirstphase.LocalandInstitutionalfunding:3.5millionUSdollars8subprojects70institutes,universities,andcompaniesinvolvedLong-termsupportasanationalproject(2006~2020)目前七十四页\总数一百一十三页\编于十五点CelebrationofCNHLPPOfficialLaunching(BeijingInstituteofRadiationMedicine)目前七十五页\总数一百一十三页\编于十五点KeylabsBeijingInstituteofRadiationMedicine目前七十六页\总数一百一十三页\编于十五点Consultingcommittee(5)ORFeome&PPISubproject,ZeguangHanChairFuchuHeHeadquartersBPRCModificationProfilesubproject,YunCaiExpressionProfileSubproject,XiaohongQianStructuralBiologySubproject,WeiminGongBioinformaticssubproject,YixueLiNewTechnologysubproject,YukuiZhangAntibodysubproject,Qi-HongSunLocalizationMappingSubproject,XueminZhang目前七十七页\总数一百一十三页\编于十五点Morethan10workshopshavebeenheld目前七十八页\总数一百一十三页\编于十五点Briefprogressofsomesubprojects

ProteinmodificationProteinlocalizationProteinstructureNewproteomicstechnologies目前七十九页\总数一百一十三页\编于十五点AnalysisofproteinphosphorylationbycombinationofIMAC,PhosphatasewithBiologicalMassSpectrometrym/zPeptidemixtureProteinsCandidatephosphopeptidesIdentificationofphosphorylatedsitesDIGESTIMACMALDI-TOFMSAnalysisPhosphatasedigestMS/MSAnalysisandDatabaseSearchingm/zPhosphopeptideConfirmationm/zMetastableionsofphosphopeptides目前八十页\总数一百一十三页\编于十五点O60716

GSLApSLDpSLRKP15924GLPSPYNMSpSAPGpSRO75379NLLEDDpSDEEEDFFLRRApSpSKGGGGYTC*QSGSGWDEFTKO95210HSpSWGDVGVGGSLKP16157RDpSRDVDEEKELLDFVPKP02671ADpSGEGDFLAEGGGVRP35221TPEELDDpSDFETEDFDVRP02730

YQpSSPAKPDSSFYKQ13813WRpSLQQLAEERYQSSPAKPDSSFpYKQ15019IYHLPDAEpSDEDEDFKEQTRYQSSPAKPDSpSFYKQ16828SNIpSPNFNFMGQLLDFERRYQSSPAKPDpSSFYKQ99959RLEISPDpSpSPERAHYTHSDYQYSQRP04792GPpSWDPFRLPEEWSQWLGGSSWPGYVRPLPPAAIEpSPAVAAPAYSRQ9H3Z4SLpSTSGESLYHVLGLDKTr:O95810SLEETLHTVDLpSpSDDDLPHDEEALEDpSAEEKVEESRP05386KEEpSEEpSDDDM*GFGLFDKEEpSEEpSDDDMGFGLFDTr:Q8NEN5pSQpSLPTTLLSPVRP08559YHGHpSMSDPGVSYRTr:Q9HAP4ESEALPEKEGEELGEGERPEEDAAALELpSpSDEAVEVEEVIEESRP11277TpSPVSLWSRLpSpSSWESLQPEPSHPYTr:Q9NQA7WLDEpSDAEMELRIdentifiedphosphopeptidesequencesfromHFL目前八十一页\总数一百一十三页\编于十五点Proteinisolationandpurificationbylectin(Glycosylation)目前八十二页\总数一百一十三页\编于十五点Co-localizationandlocomotionoftheGFPfusionproteinandDsRedDsRed-MemGreenmergeGFPGFP-fusedPrTNF-treatedGFP-fusedmutantTNF-treatedGFP-fusedPr目前八十三页\总数一百一十三页\编于十五点StructuralproteomicsCloned900genesfromliverExpressedproductsofmorethan100genesDeterminedthestructuresof3proteinsbyX-rayDeterminedthestructuresof2proteinsbyNMR目前八十四页\总数一百一十三页\编于十五点Diphosphoglyceratemutase(DPGM)X-rayProgrammedcelldeath5(PDCD5)NMR目前八十五页\总数一百一十三页\编于十五点Beijing(National)ProteomeResearchCenterChineseProteomeDevelopmentCenter目前八十六页\总数一百一十三页\编于十五点TheAntibodyBankofHumanLiverProteomeProjectEstablishment,characterization,andapplicationofantibodiesagainsthumanliverproteins

Qi-HongSunBeijingInstituteofRadiationMedicine(BIRM)BeijingProteomeResearchCenter(BPRC)目前八十七页\总数一百一十三页\编于十五点AntigensMyelomacellsHybridomacellsmAbsDiagnosticsTherapeuticsProteomicsResearch

ProfilingIdentificationQuantificationLocalizationModificationInteractionFunctionLargeantibodycollectionforproteomics目前八十八页\总数一百一十三页\编于十五点AntibodyDatabaseAntibodyBankSamples5000liverproteinsHLPPAntibodyBankAntibodychips目前八十九页\总数一百一十三页\编于十五点10%ofHLPPAntibodyBankStandardsanddatabaseofHUPOHLPP/HAIMicroarrayandotherantibody-basedtechnology90%10%HLPPAntibodyBankatPilotPhase目前九十页\总数一百一十三页\编于十五点Antigenpreparation

Fractionatedliverproteins-Unknown/native/multi-antigensRecombinantproteinsSynthesizedpeptidesAntibodygenerationHybridomacelllines–mAbs(murine/rabbit)Polyclonalantibodies–IgG(Rabbit)/orIgY(chicken)AntibodycharacterizationandapplicationClassandsubclass,ELISA,IH,WB,andIPAntibodymicroarrayandotherantibody-basedtechnologyGeneralApproach目前九十一页\总数一百一十三页\编于十五点AdultFetalDiseasedIHcharacterizationofmAbsagainsthumanliverproteins目前九十二页\总数一百一十三页\编于十五点ProgresssummaryofHLPPAntibodyBankingProject

AntigenpreparationforimmunizationFractionatedliverproteins–nucleus,membrane,microsome,mitochondrial,cytosolic,andplasmaproteinsPurifiedrecombinantproteinantigens–215Synthesizedpeptidesfromliver-specificproteins–44AntibodypreparationandcharacterizationEstablishedhybridomacelllines–1085CharacterizedmAbs–862formorethan100differentproteinsRabbitpAbs–50AntibodyApplication

Depletionofhigh-abundantproteinsImmunohistochemistryIsolationofproteincomplexes目前九十三页\总数一百一十三页\编于十五点LabsofHLPPAntibodyBankingProjectNational(China):Qi-HongSun/JianenGao,MingLi,MinZhao,DalingZhu,ChaonengJi,GangCheng,ZhinanChen,BoquanJing,JianWang/LinWu,XiaoningWang/YingLin,ZhengLi,JinliangYang,andJieTang.Internationalcollaboration:

HUPO-HAI(Dr.Uhlen)FHCRC/NCI-IBDC(Dr.Lee)GermanSystemsBiologyProject(Drs.Meyer,Ueffing)目前九十四页\总数一百一十三页\编于十五点Workingplan

Antibodydatasheet–ElectronicsubmissionformAntibodylist–Website/HLPPAntibodydatabaseAntibodydemands/applicationsAntibodyqualitycontrolAntibodyexchange,collection,anddistributionScaleofantibodybankAntibody-basedtechnologyInternationalcollaboration目前九十五页\总数一百一十三页\编于十五点目前九十六页\总数一百一十三页\编于十五点OutlineBackgroundProgressoftheHLPPInitiationoftheCNHLPPNextwork目前九十七页\总数一百一十三页\编于十五点ResearchprojectsDistributetheChinesespecimenstoreferencelabsoutsideChinaEstablishtheSOPsfortheblackracialpeople`sliversampleComprehensivelyanalyzethedataoftheproteinsexpressingprofileoftheFrenchspecimensEstablishtheSOPsfortheprotein-proteininteractionsandORFeomeInitiatevirtuallydiseasedliverproteomesubprojectInitiatevirtuallyproteomicmodificationandlocalizationsubprojects目前九十八页\总数一百一十三页\编于十五点CoordinationContinuetodiscussthecooperationwithotherinitiatives(e.g.MRPP,HPPP,PSI,HAI,etc.)orotherprogramme(e.g.SystemsBiologyProjectofGermany)EstablishtheheadquartersoftheHLPPContinuetopropagandizetheHLPPtothepublicandpriva