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ID:48737873
大小:2.39 MB
页数:28页
时间:2020-01-21
《labrlfree非标记定量蛋白质组学的分析方法.pptx》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、IntroductionLabelFreeQuantitationinMaxQuantGlyGlyDiagnosticPeaksNewAnnotationVisualizationOptionsLabelFreeQuantitation(LFQ)Comparequantitativemeasuresofproteins/peptidesBETWEENmassspecrunstoidentifysignificantchangesComparisonisWITHINthemassspecrunforSILAC,iTRAQ,andIPTLNoche
2、micalormetaboliclabelingrequiredCompatiblewithvirtuallyanysampleSpectralCounting#MS/MSidentifiedforeachproteinVerycrudeEasilyimplementedSomewhatsampledependentSpectralcountsaresometimesnormalizedacrossmassspecrunsExtractedIonChromatogram(XIC)SumofareasunderthecurvesofMS1chro
3、matogramsofeachpeptideidentifiedRequiresadvancedprocessingsoftwareSomewhatindependentofthesampleLabelFreeQuantitationinMaxQuantSILACratiocalculationrequiresthecomparisonofXICsbetweenthelabelstatesinamassspecrunCanusesameprocessingtocalculateXICsofpeptidesineachrunandthencomp
4、areacrossmassspecrunsNeedtomapfromonemassspecruntoothersEasyifMS2identifiedinbothrunsHarderifnotidentifiedinbothrunsMaxQuanthasoptiontomatchidentifiedpeakstounidentifiedpeakusingretentiontimeandaccuratemassLFQOptionsinMaxQuantPeptideLevelRawintensityonlyoptionProteinLevelRaw
5、intensityMS2countsLFQvalueNopublicationyetCalculateglobalrawfilenormalizationfactoriBAQNormalizeintensitiesbynumberofobservablepeptidesBenchmarkingLFQOptionsTriplicateyeastanalysesPearsonCorrelationTypicalcorrelationmeasureSensitivetooutliersSpearmanRankCorrelationOnlylooksa
6、trankingInsensitivetooutliersRatio95%ConfidenceIntervalTellsusthetypicalrangeofratioswewouldexpecttoseeforvaluesthatareactuallyunchangedPeptideLFQPerformanceProteinRawIntensityLFQPerformanceProteinSpectralCountsLFQPerformanceProteiniBAQLFQPerformanceProteinLFQPerformanceLFQS
7、ummaryNowpossibletogetreasonableestimatesofrelativeabundancesbetweensamplesatboththepeptideandproteinlevelProtein95%CIat~1.5foldchangefor1:1Peptide95%CIat~3foldchangefor1:1MightconsiderusingthesetechniquestomeasurelargechangesinmodificationstatusForexample,proteinfootprintin
8、gUbiquitinModificationDataMiningUbiquitinmodificationsresultinadiGlycinemod
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