人類肝臟蛋白質(zhì)組計(jì)劃的實(shí)施與毒理學(xué)發(fā)展的機(jī)遇

國(guó)際肝臟蛋白質(zhì)組學(xué)研究

方案的實(shí)施與毒理學(xué)開(kāi)展機(jī)遇1編輯ppt報(bào)告提要蛋白質(zhì)組學(xué)產(chǎn)生的時(shí)代背景“人類蛋白質(zhì)組方案〞的目標(biāo)與意義蛋白質(zhì)組學(xué)與毒理學(xué)/環(huán)境醫(yī)學(xué)2編輯ppt曼哈頓原子彈研制方案人類基因組方案阿波羅登月方案人類歷史上的三大科技工程1941.12.6—1945.7.16羅斯福批準(zhǔn),耗資20億美元原子半徑10-10m原子體積10-30m3人體半徑100m人體體積100m3太陽(yáng)系半徑1012m太陽(yáng)系體積1034m31990.10.1-2003.4.23克林頓、布萊爾批準(zhǔn)，耗資30億美元1961.5.25—1969.7.20肯尼迪批準(zhǔn),耗資240億美元3編輯ppt人類基因組方案開(kāi)創(chuàng)了“基因組時(shí)代〞4編輯ppt基因組學(xué)向蛋白質(zhì)組學(xué)“求助〞!Nature409:747,15Feb.2001Andnowforproteome

“現(xiàn)在輪到蛋白質(zhì)組〞Science291:1221,16Feb.2001ProteomicsinGenomeland“基因組大地中的蛋白質(zhì)組學(xué)〞5編輯pptPROTEIN6編輯ppt轉(zhuǎn)錄組一種細(xì)胞、組織或生物體所對(duì)應(yīng)的全套mRNARNA基因組生物體所擁有的全套染色體上的全部基因DNA蛋白質(zhì)組一種細(xì)胞、組織或生物體所對(duì)應(yīng)的全套蛋白質(zhì)PROTEIN功能執(zhí)行體遺傳信息載體7編輯ppt

基因和蛋白質(zhì)并不存在嚴(yán)格的線性關(guān)系

ORF并不預(yù)示一定存在相對(duì)應(yīng)的功能性基因

mRNA水平并非與蛋白質(zhì)的表達(dá)水平對(duì)應(yīng)翻譯后修飾及同工蛋白質(zhì)(Isoforms)等現(xiàn)象在基因水平無(wú)從表現(xiàn)蛋白質(zhì)與蛋白質(zhì)的相互作用難以在基因水平得以認(rèn)識(shí)基因組與轉(zhuǎn)錄組不能取代蛋白質(zhì)組8編輯ppt翻譯后修飾相互作用構(gòu)象變化翻譯后拼接移位胞質(zhì)胞核蛋白質(zhì)調(diào)節(jié)的多樣性生命的“萬(wàn)花筒”9編輯ppt蛋白質(zhì)功能的群體性10編輯ppt繩墻扇茅蛇樹(shù)蛋白質(zhì)作用的整體性11編輯ppt探索生命奧秘的直接對(duì)象蛋白質(zhì)組

生命體的統(tǒng)一性源于基因組生命體的多樣性、復(fù)雜性、功能性、表型源于蛋白質(zhì)組1463-4x1043x109103(1012)

12編輯ppt

Proteomicsseekstoprovidefunctionalinformationforallproteins.proteomicsismoreofaconceptthanadefinedtechnology,anditreferstoanyprotein-basedapproachthathasthecapacitytoprovidenewinformationaboutproteins

onagenomewidescale.“Proteomicsincludesnotonlytheidentificationand

quantificationofproteinsbutalsothedeterminationof

their:

localization

modifications

interactions

activities

function13編輯ppt報(bào)告提要蛋白質(zhì)組學(xué)產(chǎn)生的時(shí)代背景“國(guó)際人類肝臟蛋白質(zhì)組方案〞的目標(biāo)與意義蛋白質(zhì)組學(xué)與毒理學(xué)/環(huán)境醫(yī)學(xué)14編輯pptDNA序列圖≠基因圖人類基因組中1/3以上基因未曾確認(rèn)基因確認(rèn)的根本層次--蛋白質(zhì)水平天書(shū)解讀天書(shū)Science291:1221,200115編輯ppt全面揭示重大疾病發(fā)生發(fā)展機(jī)制的基礎(chǔ)蛋白質(zhì)組單基因病—遺傳病多基因病—腫瘤等重大疾病發(fā)生開(kāi)展機(jī)制單一基因單一蛋白基因群蛋白質(zhì)群轉(zhuǎn)錄組蛋白質(zhì)組????16編輯ppt發(fā)現(xiàn)大量新型藥靶與新藥的源泉蛋白質(zhì)組最重要的疾病100-150每種疾病相關(guān)的基因10每個(gè)基因相關(guān)的蛋白質(zhì)3–10藥靶蛋白總數(shù)

3000-15,000

現(xiàn)有藥物約2000種85％是針對(duì)目前已知的500種藥靶6-30倍藥靶有待發(fā)現(xiàn)17編輯ppt啟動(dòng)人類蛋白質(zhì)組方案勢(shì)在必行！探索人體生命奧秘的直接對(duì)象全面揭示重大疾病發(fā)生開(kāi)展機(jī)制的根底發(fā)現(xiàn)大量新型藥靶與新藥的源泉

人類基因組序列及基因注釋人類蛋白質(zhì)組VIP:VeryImportantProteome18編輯ppt人類蛋白質(zhì)組方案的

主要科學(xué)目標(biāo)驗(yàn)證人類基因組方案推測(cè)的基因，注釋基因組說(shuō)明蛋白質(zhì)組的調(diào)控模式并與轉(zhuǎn)錄組進(jìn)行比照建立蛋白質(zhì)群/組裝體，蛋白質(zhì)復(fù)合物或蛋白質(zhì)機(jī)器,即連鎖圖建立人體生理學(xué)、病理學(xué)的蛋白質(zhì)組根底19編輯ppt1463-4x1043x109103(1012)

基因組方案蛋白質(zhì)組方案20編輯ppt21編輯pptBodyfluidmostaccessibleforbiomarkerdiscoveryinanimalandhumans.LiverKidneyOrganBodyFluidBrainImmuneOrgans/VesselsIntestineOtherTissuesCSFFecesEndocrine/ParacrineSecretionsBileUrineAir/BALFLymphLungsEyesSkinOralCavitySweatSalivaTearsVenousBloodArterialBloodSerum10%90%SerumProteinsNon-InformativeAbundantProteinsi.e.albumin,IgG,etc.InformativeLowAbundanceProteinsEnrichmentStrategiesforInformativeSerumProteins:SELDI

SerumImmunosubtraction

22編輯ppt人類肝臟蛋白質(zhì)組方案里程碑蛋白表達(dá)譜蛋白連鎖圖亞細(xì)胞定位圖修飾譜人類基因組方案

里程碑遺傳圖物理圖序列圖23編輯pptInitiativesofHumanProteomeProject(HPP)HPPPHumanPlasmaProteomeProject,USAHLPPHumanLiverProteomeProject,ChinaPSIProteomicsStandardsInitiativeUKHBPP

HumanBrainProteomeProject,GermanyHAIHumanAntibodyInitiativeSweden24編輯pptWhyisliver?wwHLPP.HUPO為何研究肝臟?25編輯ppt肝臟功能的多樣性與重要性能量——能量轉(zhuǎn)換、儲(chǔ)存物質(zhì)——代謝〔活性分子的合成、毒性分子的分解〕信息——信息分子的合成與分泌26編輯ppt肝臟為其它組織提供能量—可氧化底物組織的活力依賴于高能鍵的連續(xù)生成。肝臟產(chǎn)生大局部脂肪酸，后者是禁食狀態(tài)下能量的根本來(lái)源。肝臟是給食狀態(tài)下由過(guò)剩糖合成脂肪酸的主要部位。27編輯ppt藥物毒物營(yíng)養(yǎng)物生物異源物生物轉(zhuǎn)化28編輯ppt化學(xué)多樣性向體內(nèi)的生物轉(zhuǎn)化體系提出嚴(yán)峻挑戰(zhàn)1.2×107種以上的化學(xué)物(CAService’slist)以每周約8000種的速率增加常用化學(xué)物在63,000種以上大約11,500種，作為食物或藥物制劑添加物攝入其余的50,000種，為潛在的環(huán)境污染物29編輯ppt人類發(fā)育過(guò)程中造血組織的興替造血系統(tǒng)的發(fā)育必需肝臟的“培育〞30編輯ppt合成大多數(shù)循環(huán)血漿蛋白合成和分泌血漿蛋白是肝臟實(shí)質(zhì)細(xì)胞——肝細(xì)胞的獨(dú)有功能〔肝細(xì)胞占肝臟總細(xì)胞數(shù)的60%及肝臟質(zhì)量的約80%〕最有特征的血漿蛋白是白蛋白，在大多數(shù)哺乳動(dòng)物中占總血漿蛋白的55-60%凝血酶、抗凝因子、溶栓酶等31編輯ppt肝臟再生機(jī)體再生能力最強(qiáng)的器官終生保持旺盛的再生能力32編輯ppt肝臟中包含的“組〞(-ome)Metabolicgenomics→Metabolome

(代謝組)Energygenomics→Energome

(能量組)Pharmacogenomics→Pharmacome

(藥理組)Toxicogenomics→Toxicome(毒理組)Regeneratinggenomics

→Regenerome

(再生組)33編輯ppt全球及中國(guó)肝炎的流行病學(xué)統(tǒng)計(jì)

全球:3.5億攜帶者中國(guó):1.8億攜帶者死亡:23萬(wàn)人/年疾病負(fù)擔(dān):500億元人民幣治療手段:抗病毒,保肝降 酶,抗纖維化,免疫治療等缺點(diǎn):病毒易反彈,病 情易反復(fù)34編輯ppt肝炎向肝癌的惡性轉(zhuǎn)化

難以遏制全世界現(xiàn)有3.5億慢性乙肝病毒〔HBV〕攜帶者，占世界人口的5%，亞洲和非洲HBV攜帶率為8-15%HBV攜帶者中50-70%病毒復(fù)制活潑，為慢性乙肝慢性乙肝病人肝硬化發(fā)生率為2-20%，代償性肝硬化開(kāi)展成失代償性肝硬化為20-23%，開(kāi)展成肝癌的占6-15%中國(guó)的慢性乙肝病人中，約25-40%最終將死于肝硬化或合并肝癌HBV攜帶者最終死于相關(guān)肝病的危險(xiǎn)性，男性為50%，女性為15%35編輯ppt全球:100萬(wàn)人/年中國(guó):50萬(wàn)人/年死亡:約45萬(wàn)人/年疾病負(fù)擔(dān):400億元人民幣治療手段:手術(shù)切除(只有 15%-20%)治療效果:術(shù)后易轉(zhuǎn)移,易復(fù) 發(fā),五年存活率 40-50%全球及中國(guó)肝癌的流行病學(xué)統(tǒng)計(jì)36編輯ppt原發(fā)性肝癌的治療水平亟待突破世界上每年有100萬(wàn)新發(fā)原發(fā)性肝癌病人，其中40-50%在我國(guó)

我國(guó)原發(fā)性肝癌發(fā)病率和死亡率均位居第二位

近20年來(lái)我國(guó)肝癌的發(fā)病率上升近40%原發(fā)性肝癌一般起病較隱匿，早期缺乏典型臨床表現(xiàn)，初診大多已屬中晚期

初診肝癌病人中，只有15-20%的病人具備手術(shù)條件這些病人術(shù)后5年生存率僅為40-50%

肝癌是致死率最高的惡性腫瘤之一37編輯ppt重大肝病的發(fā)生開(kāi)展無(wú)法歸咎于少數(shù)幾個(gè)基因或蛋白質(zhì)所呈現(xiàn)的功能狀態(tài)和信號(hào)通路從蛋白質(zhì)組層面上“全景式〞揭示肝臟疾病的生理病理機(jī)制是解決重大肝病的根本出路肝炎病毒肝臟/肝細(xì)胞肝炎肝硬化/纖維化原發(fā)肝癌肝癌轉(zhuǎn)移多因素、多步驟的發(fā)病機(jī)制38編輯ppt肝臟是人類蛋白質(zhì)組方案的首批目標(biāo)！39編輯pptPrometheusstolefirefromthegodsandgaveittomortals.PrometheusBoundHeraclesLiberatePrometheusHLPP—ModernPrometheusMythPrometheus

LiverEagleLiverdiseases

HeraclesHLPP40編輯ppt人類肝臟蛋白質(zhì)組方案

HumanLiverProteomeProject(HLPP)國(guó)際HLPP共同體41編輯pptGenerateanintegrativeapproachleadingtoacomprehensivefunctionalmapoftheliver

Expandliverproteometoits“PHYSIOME〞and“PATHOME〞todramaticallyacceleratethedevelopmentofdiagnostics,preventionandtherapeuticstowardsitsdiseases

Developstandardoperatingprocedures(SOPs)forotherHUPOInitiativesVISION42編輯pptHUPOWorkshop,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.,2005MISION43編輯ppt表達(dá)譜修飾譜定位圖連鎖圖樣品庫(kù)抗體庫(kù)數(shù)據(jù)庫(kù)HLPP的科學(xué)目標(biāo)---肝臟蛋白質(zhì)組的“太陽(yáng)系〞3-D圖ORF組44編輯ppt肝臟蛋白質(zhì)組與

肝臟生理、病理的銜接肝臟蛋白質(zhì)組代謝組毒理組藥理組再生組肝炎組肝癌組能量組45編輯ppt作為分泌器官，肝臟合成大多數(shù)的循環(huán)血漿蛋白

肝臟轉(zhuǎn)錄組“肝臟蛋白質(zhì)組方案〞和“血漿蛋白質(zhì)組方案〞的整合肝臟蛋白質(zhì)組血漿蛋白質(zhì)組?46編輯ppt基因組肝臟蛋白質(zhì)組肝臟轉(zhuǎn)錄組基因組、轉(zhuǎn)錄組和蛋白質(zhì)組的集成證實(shí)推測(cè)基因比照調(diào)控模式47編輯ppt重要功能蛋白質(zhì)肝病藥物靶標(biāo)肝病診斷標(biāo)志物人類肝臟蛋白質(zhì)組48編輯ppt肝炎病毒肝臟/肝細(xì)胞感染肝炎肝硬化/纖維化原發(fā)肝癌肝癌轉(zhuǎn)移重大肝病預(yù)警、診斷、預(yù)防、治療的關(guān)鍵環(huán)節(jié)“東亞病夫〞“肝病大國(guó)〞1億多肝炎病毒攜帶者1000億多/年醫(yī)療費(fèi)用49編輯pptSOPsSpecimenBankingPlatformsWorkingteamGlobalCollaborationExpressionProfileORFeomeAntibodiesBioinformatics…

ModificationProfileSubcellularLocalizationProtein-ProteinInteractionAntibodiesBioinformaticsIntegrateproteomewithtranscriptomeCompareproteomeswithinhealthanddiseasedliverTimeLinesPilotPhase(2003-2005)PhaseII(2006-2021)50編輯pptOutlineBackgroundProgressoftheHLPP-ProgressreportonexpressionprofilingInitiationoftheCNHLPPNextwork51編輯pptProgress-ExpressionprofilingEvaluationoftechnologiesBioinformaticsset-upSamplingandpreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues52編輯pptProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues53編輯pptMajorconclusionsfromthevaluationoftechnologies–16standardproteins/7labsHighmolecularweightandlowabundanceproteinsarelessdetectableby2DE.Proteinswithonlythreeorderofmagnitudecouldbeidentifiedin2DE,butproteinswithfourorderofmagnitudecouldbeidentifiedinshotguntechnology.Morepeptides(redundant)wereidentifiedforhighMWproteinsinshotguntechnology.Somesimilarproteinswereidentified(notproteingroup),suggestingthecriteriafordistinguishingsimilarproteinsshouldbemorestrict.54編輯pptProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues55編輯pptHLPPExperimentalDatabaseMWSDataClient…GUI，WebServices,APIFilesInterchange,DMBSDumpDatasynchronizeDatasubmissionExperimentreports,data,files…

HLPPDataMirrorMWSDataClientMWSDataClientMWSDataClientMWSDataClientMWSServerAdministratorHLPPDataMirrorSubmissionPackagesRepositoryHLPPProjectManagementDatabaseHLPPProjectManagementSystemHLPPParticipatingLaboratoriesHLPPDataCenterTheHLPPDataManagementSystem

ArchitectureBasedonMyWorkSpaceSystem(MWS)56編輯pptCurrentStandarddraftofHLPPSampleInformationProteinextractionConcentrationMeasurementSeparationGel-based2DEGel1D-IEFGel1D-SDSLC-basedRPLCSCXLCSAXLCSECTrapDesaltingDigestionIonsourceMALDIAutoflexUltraflexABI4700ESIQstarQTofMicroLCQLTQMassspectrometryTOFTOFTOFUltraflexABI4700QTOFQstarQTofMicroITMSLCQLTQPeaklistgenerationABI4700QStarAutoflexTurboSequestMasslynxFlexAnalysisPeaklistpreprocessingGPSpeaklistpreprocessProtein/peptideidentificationMascotSequestProteinlistgenerationAutoflexGPSBiotoolsQstarBuildSummaryDTASelectBioworksAutoQuest57編輯pptProteinidentificationisbasedontheanalysisofpeptidesgeneratedbyproteolyicdigestion.Whenthedetectedpeptidesmatchmanyproteinsandcannotidentifyauniqueprotein,theresultwillbepresentedintheformofagroupofpossibleproteins.

DefinitionofProteinGROUP

58編輯pptCriteriaforproteinidentificationIontrapXCorr(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)59編輯pptProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandpreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues60編輯pptEvaluationoftheSOPsforthesubcellularfractionationObject:FindtheoptimummethodofpretreatmentSample:LivertissuesofC57miceSubcellular:Mitochondria,Golgi,PM,Nucleus,ER,CytoplasmSamplepretreatmentFreshtissuesFrozenhomogenisedtissuesFrozentissues61編輯pptPurity(Westernblotting):Freshtissues>frozenhomogenizedtissues>frozentissuesTheyieldofproteins:Freshtissues>frozenhomogenizedtissues>frozentissuesIntegrity(TEM):Freshtissues>frozenhomogenizedtissuesSummary62編輯pptProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues63編輯pptItisagiantambitiousobjectiveandgivesagreatchallengetoembryonicproteomicsItisnecessarytocarryoutapreviewbeforetheformallaunchingofthisprojectScience302:1316,Nov.21,2003Nature425:441,Oct.2,200364編輯pptAverageThroughputofthePlatform2DESystem

12Gel/3DaysAuto-SpotDigestSystem

1152spots/DayMSandMS/MSSystem

500-1000Spots/Day65編輯pptTheflowchartofCCPITfortheproteinexpressionprofileofhumanfetalliver66編輯pptTheareascalesbeforeandafterisoformprocessingingroupandproteinlevelsrespectively.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:

125867編輯pptACBChromosomaldistributionofHFLproteome-encodinggenes68編輯pptFunctionalModulesinplasmamembrane69編輯pptProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues70編輯ppt

ReferenceLabsforHLPPExpressionProfilingProject

FuchuHe&XiaohongQian(BeijingInstituteofRadiationMedicine)RongZeng(ShanghaiInstituteforBiologicalSciences)PengyuanYang(FudanUniversity)SiqiLiu(BeijingGenomicsInstitute,ChineseAcademyofSciences)71編輯pptLauraBeretta(FHCRC,USA)RichardJ.Simpson(RoyalMelbourneHospital,Australia)

AngelikaGorg(TechnischeUniversitatMunchen,Germany)MarkBaker/JunhongSun(APAF,Australia)72編輯pptOutlineBackgroundProgressoftheHLPPInitiationoftheCNHLPPNextwork73編輯pptCNHLPPOverviewThefirstphase:2004-2005Fundingfromcentralgovernment:10millionUSdollars ChineseMOST:jointlyfundedbyNationalProgramon KeyBasicResearchProjects(973),NationalHigh-tech R&DProgram(863),andNationalKeyTechnologiesR&D Programmeininthefirstphase.LocalandInstitutionalfunding:3.5millionUSdollars8subprojects70institutes,universities,andcompaniesinvolvedLong-termsupportasanationalproject(2006~2021)74編輯pptCelebrationofCNHLPPOfficialLaunching(BeijingInstituteofRadiationMedicine)75編輯pptKeylabsBeijingInstituteofRadiationMedicine76編輯pptConsultingcommittee(5)ORFeome&PPISubproject,ZeguangHanChairFuchuHeHeadquartersBPRCModificationProfilesubproject,YunCaiExpressionProfileSubproject,XiaohongQianStructuralBiologySubproject,WeiminGongBioinformaticssubproject,YixueLiNewTechnologysubproject,YukuiZhangAntibodysubproject,Qi-HongSunLocalizationMappingSubproject,XueminZhang77編輯pptMorethan10workshopshavebeenheld78編輯pptBriefprogressofsomesubprojects

ProteinmodificationProteinlocalizationProteinstructureNewproteomicstechnologies79編輯pptAnalysisofproteinphosphorylationbycombinationofIMAC,PhosphatasewithBiologicalMassSpectrometrym/zPeptidemixtureProteinsCandidatephosphopeptidesIdentificationofphosphorylatedsitesDIGESTIMACMALDI-TOFMSAnalysisPhosphatasedigestMS/MSAnalysisandDatabaseSearchingm/zPhosphopeptideConfirmationm/zMetastableionsofphosphopeptides80編輯pptO60716

GSLApSLDpSLRKP15924GLPSPYNMSpSAPGpSRO75379NLLEDDpSDEEEDFFLRRApSpSKGGGGYTC*QSGSGWDEFTKO95210HSpSWGDVGVGGSLKP16157RDpSRDVDEEKELLDFVPKP02671ADpSGEGDFLAEGGGVRP35221TPEELDDpSDFETEDFDVRP02730

YQpSSPAKPDSSFYKQ13813WRpSLQQLAEERYQSSPAKPDSSFpYKQ15019IYHLPDAEpSDEDEDFKEQTRYQSSPAKPDSpSFYKQ16828SNIpSPNFNFMGQLLDFERRYQSSPAKPDpSSFYKQ99959RLEISPDpSpSPERAHYTHSDYQYSQRP04792GPpSWDPFRLPEEWSQWLGGSSWPGYVRPLPPAAIEpSPAVAAPAYSRQ9H3Z4SLpSTSGESLYHVLGLDKTr:O95810SLEETLHTVDLpSpSDDDLPHDEEALEDpSAEEKVEESRP05386KEEpSEEpSDDDM*GFGLFDKEEpSEEpSDDDMGFGLFDTr:Q8NEN5pSQpSLPTTLLSPVRP08559YHGHpSMSDPGVSYRTr:Q9HAP4ESEALPEKEGEELGEGERPEEDAAALELpSpSDEAVEVEEVIEESRP11277TpSPVSLWSRLpSpSSWESLQPEPSHPYTr:Q9NQA7WLDEpSDAEMELRIdentifiedphosphopeptidesequencesfromHFL81編輯pptProteinisolationandpurificationbylectin(Glycosylation)82編輯pptCo-localizationandlocomotionoftheGFPfusionproteinandDsRedDsRed-MemGreenmergeGFPGFP-fusedPrTNF-

treatedGFP-fusedmutantTNF-

treatedGFP-fusedPr83編輯pptStructuralproteomicsCloned900genesfromliverExpressedproductsofmorethan100genesDeterminedthestructuresof3proteinsbyX-rayDeterminedthestructuresof2proteinsbyNMR84編輯pptDiphosphoglyceratemutase(DPGM)X-rayProgrammedcelldeath5(PDCD5)NMR85編輯pptBeijing(National)ProteomeResearchCenterChineseProteomeDevelopmentCenter86編輯pptTheAntibodyBankofHumanLiverProteomeProjectEstablishment,characterization,andapplicationofantibodiesagainsthumanliverproteins

Qi-HongSunBeijingInstituteofRadiationMedicine(BIRM)BeijingProteomeResearchCenter(BPRC)87編輯pptAntigensMyelomacellsHybridomacellsmAbsDiagnosticsTherapeuticsProteomicsResearch

ProfilingIdentificationQuantificationLocalizationModificationInteractionFunctionLargeantibodycollectionforproteomics88編輯pptAntibodyDatabaseAntibodyBankSamples5000liverproteinsHLPPAntibodyBankAntibodychips89編輯ppt10%ofHLPPAntibodyBankStandardsanddatabaseofHUPOHLPP/HAIMicroarrayandotherantibody-basedtechnology90%10%HLPPAntibodyBankatPilotPhase90編輯pptAntigenpreparation

Fractionatedliverproteins-Unknown/native/multi-antigensRecombinantproteinsSynthesizedpeptidesAntibodygenerationHybridomacelllines–mAbs(murine/rabbit)Polyclonalantibodies–IgG(Rabbit)/orIgY(chicken)AntibodycharacterizationandapplicationClassandsubclass,ELISA,IH,WB,andIPAntibodymicroarrayandotherantibody-basedtechnologyGeneralApproach91編輯pptAdultFetalDiseasedIHcharacterizationofmAbsagainsthumanliverproteins92編輯pptProgresssummaryofHLPPAntibodyBankingProject

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

Depletionofhigh-abundantproteinsImmunohistochemistryIsolationofproteincomplexes93編輯pptLabsofHLPPAntibodyBankingProjectNational(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)94編輯pptWorkingplan

Antibodydatasheet–ElectronicsubmissionformAntibodylist–Website/HLPPAntibodydatabaseAntibodydemands/applicationsAntibodyqualitycontrolAntibodyexchange,collection,anddistributionScaleofantibodybankAntibody-basedtechnologyInternationalcollaboration95編輯ppt96編輯pptOutlineBackgroundProgressoftheHLPPInitiationoftheCNHLPPNextwork97編輯pptResearchprojectsDistributetheChinesespecimenstoreferencelabsoutsideChinaEstablishtheSOPsfortheblackracialpeople`sliversampleComprehensivelyanalyzethedataoftheproteinsexpressingprofileoftheFrenchspecimensEstablishtheSOPsfortheprotein-proteininteractionsandORFeomeInitiatevirtuallydiseasedliverproteomesubprojectInitiatevirtuallyproteomicmodificationandlocalizationsubprojects98編輯pptCoordinationContinuetodiscussthecooperationwithotherinitiatives(e.g.MRPP,HPPP,PSI,HAI,etc.)orotherprogramme(e.g.SystemsBiologyProjectofGermany)EstablishtheheadquartersoftheHLPPContinuetopropagandizetheHLPPtothepublicandprivatedomainsStriveformorefinancialsupportsfrommoreresources99編輯pptHLPPworkshopsJun.3,Washington Organizer:LauraBeretaJun.10,EBI/London Organizer:RolfApweilerAug.27,Munich Organizer:FuchuHe100編輯ppt報(bào)告提要蛋白質(zhì)組學(xué)產(chǎn)生的時(shí)代背景“人類蛋白質(zhì)組方案〞的目標(biāo)與意義蛋白質(zhì)組學(xué)與毒理學(xué)/環(huán)境醫(yī)學(xué)101編輯ppt主要作用揭示環(huán)境因素的致病機(jī)制了解化學(xué)毒物的代謝途徑解析微生物蛋白質(zhì)組組成增強(qiáng)疾病預(yù)防診斷與治療102編輯pptGenetics

Influence

Environment

GenomicsTranscriptomics2-DE/MSICAT-LC/MSInformati