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Intracellular Accumulation of Lipids via Scavenger Receptors 2021/4/261Formation of foam cellAthersclerosisisachronicinflammatoryreactioncausedbymacrophagesAthersclerosisisachronicinflammatoryreactioncausedbymacrophagesandlymphocytesthatuptakethepathogenicitylipidsinvadingthevessel.andlymphocytesthatuptakethepathogenicitylipidsinvadingthevessel.2021/4/262SR-AandCD36aretheSR-AandCD36aretheprincipalreceptorsprincipalreceptorsresponsibleforfoam-responsibleforfoam-cellformationcellformationMacrophage cell takes up lipid by SR-A2021/4/263Structure of SR-A SixdomainsSixdomains ThreetypesThreetypes TrimericTrimericmoleculemolecule2021/4/264 FoamcellformationMadhesionHostdefenseandinnateimmunityAdaptiveresponsesDisordersofthenervoussystemPhagocytosisofapoptoticcellsFunctions of SR-A 2021/4/265The cytoplasmic domain is the key part of the receptor2021/4/266 mediatetheinternalizationofmediatetheinternalizationofligandsligandssuchasox-LDLsuchasox-LDL participatetheregulationofcelladhesionandmigrationparticipatetheregulationofcelladhesionandmigration accommodatetheexpressionandtransportofreceptoraccommodatetheexpressionandtransportofreceptor link-coupledwithdownstreamofsignalingpathwaylink-coupledwithdownstreamofsignalingpathwayFunctions of SR-A cytoplasmic domain:2021/4/267 Cytoplasmic domain：MurineMurine:MTKEMTENQRLCPHEREDADCSSES:MTKEMTENQRLCPHEREDADCSSESVKFDARSVKFDARSMTAMTASLSLPPHSTKNGPSVQEHSTKNGPSVQEKLKSFKKLKSFK(55)(55)Rabbit:MAQWDSFTDQQEDTDSCSESRabbit:MAQWDSFTDQQEDTDSCSESVKFDARSVKFDARSNTANTALLLLPPNPKPPNPKGPPLQEGPPLQEKLKSFKKLKSFK(5050)Bovine:MAQWDDFPDQQEDTDSCTESBovine:MAQWDDFPDQQEDTDSCTESVKFDARSVKFDARSVTAVTALLLLPPHPKNPPHPKNGPTLQERMKSYKGPTLQERMKSYK(50)(50)Human:MEQWDHFHNQQEDTDSCSESHuman:MEQWDHFHNQQEDTDSCSESVKFDARSVKFDARSMTAMTALLLLPPNPKPPNPKNSPSLQENSPSLQEKLKSFKKLKSFK(50)(50)2021/4/268MSR-AAcLDLHSP90,HSP70,andGAPDHDirectlyInteractwiththeCytoplasmicDomainofMSRNakamuraT.etal.BBRC,290,858-864(2002)FunctionalandpossiblephysicalassociationofscavengerreceptorwithcytoplasmictyrosinekinaseLyninmonocyticTHP-l-derivedMacrophagesMikiS.etal.FEBSLetters,399,241-244(1996)HSP90 HSP70 GAPDHLyn2021/4/269LigandBindingtoMacrophageScavengerReceptor-AInducesUrokinase-typePlasminogenActivatorExpressionbyaProteinKinase-dependentSignalingPathwayHsien-YehHsuetal.JBiolChem,273,1240-1246(1998)MSR-AAcLDLFucoidanPTKPLC-PKCuPA2021/4/26109、人的价值，在招收诱惑的一瞬间被决定。2024/7/72024/7/7Sunday,July7,202410、低头要有勇气，抬头要有低气。2024/7/72024/7/72024/7/77/7/202411:04:22AM11、人总是珍惜为得到。2024/7/72024/7/72024/7/7Jul-2407-Jul-2412、人乱于心，不宽余请。2024/7/72024/7/72024/7/7Sunday,July7,202413、生气是拿别人做错的事来惩罚自己。2024/7/72024/7/72024/7/72024/7/77/7/202414、抱最大的希望，作最大的努力。07七月20242024/7/72024/7/72024/7/715、一个人炫耀什么，说明他内心缺少什么。七月242024/7/72024/7/72024/7/77/7/202416、业余生活要有意义，不要越轨。2024/7/72024/7/707July202417、一个人即使已登上顶峰，也仍要自强不息。2024/7/72024/7/72024/7/72024/7/7MSR-AFucoidanHsp90,pp60SrcRac1PAKJNKp38PI3KproIL-1IL-1LigandsofMacrophageScavengerReceptorInduceCytokineExpressionviaDifferentialModulationofProteinKinaseSignalingPathwaysHsuHY.etal.JBiolChem,276,28719-28730(2001)2021/4/2612MSR-AAcLDLinternalization,adhesionGi/oRegulationofacetylatedlowdensitylipoproteinuptakeinmacrophagesbypertussistoxin-sensitiveGproteinsStewartCetal.JLipidRes,41,807-813(2000)ClassAscavengerreceptorsmediatecelladhesionviaactivationofGi/oandformationoffocaladhesioncomplexes PostSR.etal.JLipidRes,43,1829-1836(2002)2021/4/2613 phosphorylationphosphorylationofcytoplasmicdomainofcytoplasmicdomain signalsequenceinsignalsequenceincytoplasmiccytoplasmicdomaindomain interactorsinteractorswithcytoplasmicdomainwithcytoplasmicdomainFocus on cytoplasmic domain：2021/4/2614BovineModelHumanModelMurineModelRabbitModel2021/4/2615Ser21（rat）increasesexpressionofreceptorpromotesmetabolismoflipidFongLGetal.JBiolChem,274,36088-36816(1999)phosphorylationofcytoplasmicdomain2021/4/2616phosphorylationofcytoplasmicdomainSer49（rat）reducesuptakeoflipidFongLGetal.JBiolChem,274,36088-36816(1999)2021/4/2617FDARS internalizationofreceptor FongLGetal.JBiolChem,274,36088-36816(1999)signalsequenceofcytoplasmicdomain2021/4/2618KLKSFK celladhesionKosswigNetal.JBiolChem,278,34219-34225(2003)signalsequenceofcytoplasmicdomain2021/4/2619VXFD(rabbit)internalizationofreceptor,transferencetocellsurfaceMorimotoKetal.BiolPharmBull,22,1022-1026(1999)signalsequenceofcytoplasmicdomain2021/4/2620Ser48(human）expressionandinternalization ofreceptor,cellmigration HeiderHetal.FEBSLetters,505,185-190(2001)phosphorylationofcytoplasmicdomain2021/4/2621Di-leucine motif?signalsequenceofcytoplasmicdomain2021/4/2622 1.The role of Di-leucine motif in the cytoplasmic domain of SRA2021/4/2623(1)SR-A mutants/EGFP constructs:Wild typeN3132LMEGFPSRASRA2021/4/2624(2)Expression patterns of SRA mutant:*P0.05,n=3EGFPSR-AN3132LMEGFPSR-AN3132LM*0 020020040040060060080080010001000controlcontrolEGFPEGFPSR-ASR-AN3132LMN3132LM2021/4/2625(3)The internalization of DiI-AcLDL via SRA mutant:DiI-acLDLSR-Aexpression*DiI-acLDL*P0.05,n=3*P0.05,n=30 01010202030304040EGFPEGFPWild typeWild typeN3132LMN3132LM*EGFPEGFPWild typeWild typeN3132LMN3132LM0.020.020.040.040.060.060.080.080.100.100.120.122021/4/2626(4)Cell adhesion via SRA mutant:0 00.10.10.20.20.30.30.40.40.50.50.60.60.70.70.80.80.90.9EGFPEGFPSR-A SR-A N3132LMN3132LMratioratio%*P0.05,n=3*2021/4/2627EGFPSR-AN3132LMImmunofluorescence:TheCHOcellsweretransfectedwithSRA/EGFPorSRAMutant/EGFP,(5)Di-leucine motif and Clathrin-coated pit:GFPAnti-clathrin overlay 2021/4/2628180KD97KDabca:EGFP-transfectedcellb:SR-A-transfectedcellc:N3132LMmutant-transfectedcellClathrin IP:2021/4/2629N1-27N1-27LM*P0.05,n=3SR-AN1-27N1-27LM97KDA AB BC C(6)Expressing of the SRA mutants in CHO cellEGFPEGFPSR-ASR-A40040060060080080010001000N1-27N1-27LM2002002021/4/2630(7)The internalization of DiI-AcLDL via SRA mutant:*P0.05,n=3*P3.(3)Isolation of binding phage clones by Phage ELISA:II4II5II6II7II9II10II11II12M1300.10.20.30.40.50.60.70.8二4二5二6二7二9二10二11二12M132021/4/2636(4)Sequencing the selected positive clones:*H11:TDRLFMNSIWPG(4,6,7,10,11,12)*H11:TDRLFMNSIWPG(4,6,7,10,11,12)*H5:TFWDDGWLLLPR(5)*H5:TFWDDGWLLLPR(5)*H9:YGDRWFMPVMRS(9)*H9:YGDRWFMPVMRS(9)2021/4/2637(5)Searching the PIR-PSD database:H5:CaseinkinaseIIphosphorylationsite(H5:CaseinkinaseIIphosphorylationsite(TFWDTFWD)H11:ProteinkinaseCphosphorylationsite(H11:ProteinkinaseCphosphorylationsite(TDRTDR)2021/4/2638(6)Construction of expression plasmidsPEGFP-N1-H5PEGFP-N1-H9PEGFP-N1-H112021/4/2639EGFPEGFPPeptide/EGFPPeptide/EGFP(7)Subcellular localization of the peptide/EGFP in CHO cells expressing SRA2021/4/2640(8)DiI-AcLDL uptake assayEGFP-H11inhibitsSRA-mediatedDiI-AcLDLassociation.CHOcellsstablyexpressingSRAweretransfectedwithEGFP-N1,EGFP-N1-H11,EGFP-N1-H9,thenincubatedwithDiIAcLDL(5ug/ml)for2hours.*p=0.03710.05*2021/4/2641AcknowledgementYaoyuChenXiaohuaWangYaoyuChenXiaohuaWangJingjingBenHuiBaiJingjingBenHuiBaiLemingFanYongJiLemingFanYongJiXiuyingChenXiuyingChen2021/4/2642ThanksThanks2021/4/26439、人的价值，在招收诱惑的一瞬间被决定。2024/7/72024/7/7Sunday,July7,202410、低头要有勇气，抬头要有低气。2024/7/72024/7/72024/7/77/7/202411:04:23AM11、人总是珍惜为得到。2024/7/72024/7/72024/7/7Jul-2407-Jul-2412、人乱于心，不宽余请。2024/7/72024/7/72024/7/7Sunday,July7,202413、生气是拿别人做错的事来惩罚自己。2024/7/72024/7/72024/7/72024/7/77/7/202414、抱最大的希望，作最大的努力。07七月20242024/7/72024/7/72024/7/715、一个人炫耀什么，说明他内心缺少什么。七月242024/7/72024/7/72024/7/77/7/202416、业余生活要有意义，不要越轨。2024/7/72024/7/707July202417、一个人即使已登上顶峰，也仍要自强不息。2024/7/72024/7/72024/7/72024/7/7谢谢大家谢谢大家2021/4/2644
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