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ID:39969709
大小:2.39 MB
页数:28页
时间:2019-07-16
《labrlfree非标记定量蛋白质组学的分析方法x》由会员上传分享,免费在线阅读,更多相关内容在教育资源-天天文库。
1、IntroductionLabelFreeQuantitationinMaxQuantGlyGlyDiagnosticPeaksNewAnnotationVisualizationOptionsLabelFreeQuantitation(LFQ)Comparequantitativemeasuresofproteins/peptidesBETWEENmassspecrunstoidentifysignificantchangesComparisonisWITHINthemassspecrunforSILAC,iTRAQ,andIPTLNochemicalormetaboliclabelin
2、grequiredCompatiblewithvirtuallyanysampleSpectralCounting#MS/MSidentifiedforeachproteinVerycrudeEasilyimplementedSomewhatsampledependentSpectralcountsaresometimesnormalizedacrossmassspecrunsExtractedIonChromatogram(XIC)SumofareasunderthecurvesofMS1chromatogramsofeachpeptideidentifiedRequiresadvanc
3、edprocessingsoftwareSomewhatindependentofthesampleLabelFreeQuantitationinMaxQuantSILACratiocalculationrequiresthecomparisonofXICsbetweenthelabelstatesinamassspecrunCanusesameprocessingtocalculateXICsofpeptidesineachrunandthencompareacrossmassspecrunsNeedtomapfromonemassspecruntoothersEasyifMS2iden
4、tifiedinbothrunsHarderifnotidentifiedinbothrunsMaxQuanthasoptiontomatchidentifiedpeakstounidentifiedpeakusingretentiontimeandaccuratemassLFQOptionsinMaxQuantPeptideLevelRawintensityonlyoptionProteinLevelRawintensityMS2countsLFQvalueNopublicationyetCalculateglobalrawfilenormalizationfactoriBAQNorma
5、lizeintensitiesbynumberofobservablepeptidesBenchmarkingLFQOptionsTriplicateyeastanalysesPearsonCorrelationTypicalcorrelationmeasureSensitivetooutliersSpearmanRankCorrelationOnlylooksatrankingInsensitivetooutliersRatio95%ConfidenceIntervalTellsusthetypicalrangeofratioswewouldexpecttoseeforvaluestha
6、tareactuallyunchangedPeptideLFQPerformanceProteinRawIntensityLFQPerformanceProteinSpectralCountsLFQPerformanceProteiniBAQLFQPerformanceProteinLFQPerformanceLFQSummaryNowpossibletogetreasonableestimatesofrelativeabundancesbetweensamplesatboththepeptideandproteinlevelProtein95%CIat~1.5foldchangefor1
7、:1Peptide95%CIat~3foldchangefor1:1MightconsiderusingthesetechniquestomeasurelargechangesinmodificationstatusForexample,proteinfootprintingUbiquitinModificationDataMiningUbiquitinmodificationsresultinadiGlycinemod
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