Acyl Chains of Phospholipase D Transphosphatidylation Products in Arabidopsis Cells A Study Using Multiple Reaction Monitoring Mass Spectrometry 英文参考文献.docVIP

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Acyl Chains of Phospholipase D Transphosphatidylation Products in Arabidopsis Cells A Study Using Multiple Reaction Monitoring Mass Spectrometry 英文参考文献.doc

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Acyl Chains of Phospholipase D Transphosphatidylation Products in Arabidopsis Cells A Study Using Multiple Reaction Monitoring Mass Spectrometry 英文参考文献

AcylChainsofPhospholipaseDTransphosphatidylation ProductsinArabidopsisCells:AStudyUsingMultiple ReactionMonitoringMassSpectrometry DominiqueRainteau1,LydieHumbert1,EliseDelage2,3,ChantalVergnolle2,3,CatherineCantrel2,3 ,Marie- AnneMaubert1,SandrineLanfranchi2,3,Re′gisMaldiney2,3,SylvieCollin2,3,ClaudeWolf1, AlainZachowski2,3,EricRuelland2,3 * 1UPMCUnivParis06,ERLINSERMU1057/UMR7203,Paris,France,2UPMCUnivParis06,UR5,PhysiologieCellulaireetMole′culairedesPlantes,Paris,France,3CNRS, EAC7180,Paris,France Abstract Background:PhospholipasesD(PLD)aremajorcomponentsofsignallingpathwaysinplantresponsestosomestressesand hormones.TheproductofPLDactivityisphosphatidicacid(PA).PAswithdifferentacylchainsdonothavethesameprotein targets, so to understand the signalling role of PLD it is essential to analyze the composition of its PA products in the presenceandabsenceofanelicitor. Methodology/Principalfindings:PotentialPLDsubstratesandproductswerestudiedinArabidopsisthalianasuspension cells treated with or without the hormone salicylic acid (SA). As PA can be produced by enzymes other than PLD, we analyzedphosphatidylbutanol(PBut),whichisspecificallyproducedbyPLDinthepresenceofn-butanol.Theacylchain compositionsofPButandthemajorglycerophospholipidsweredeterminedbymultiplereactionmonitoring(MRM)mass spectrometry.PButprofilesofuntreatedcellsorcellstreatedwithSAshowanover-representationof160/18:2-and16:0/ 18:3-species compared to those of phosphatidylcholine and phosphatidylethanolamine either from bulk lipid extracts or frompurifiedmembranefractions.WhenmicrosomalPLDswereusedininvitroassays,theresultingPButprofilematched exactly that of the substrate provided. Therefore there is a mismatch between the acyl chain compositions of putative substratesandtheinvivoproductsofPLDsthatisunlikelytoreflectanyselectivityofPLDsfortheacylchainsofsubstrates. Conclusions:MRMmassspectrometryisareliabletechniquetoanalyzePLDproducts.OurresultssuggestthatPLDaction inresponsetoSAisnotduetotheproductiono