Chapter 24 Targeted Proteomics

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1、CHAPTER24TargetedProteomicsINTRODUCTIONProteomicsisarelativelynewdisciplinethathasgrownrapidly,bothintermsofexpectationsandcapabilities,duringthepastdecade.Chapters22(Gel-BasedPro-teomeAnalysis)and23(Non-Gel-BasedProteomeAnalysis)focusprimarilyonprotein-profilingtechniquesand

2、associatedanalysisstrategies.Thetechniquesdescribedinthosechapterscanbeappliedtodifferenttypesofsampleshavingawiderangeofcomplexity.Comprehensive(global)proteomeanalysisinvolvesattemptstoidentifyand,insomecases,quantitativelycompareasmanyproteinsaspossibleinthesampleofintere

3、st.However,comprehensiveanalysisofcomplexproteomes,suchasthosefromcellsortissuesofhighereukaryoticorganisms,iscomplicatedbythefactthatthecomplexityoftheproteomebeinganalyzedusuallygreatlyexceedstheanalyticalcapacityofcommoncurrentprotein-profilingmethodssuchas2-DgelsorLC/LC-M

4、S/MSanalysis.Ausefulstrategythatfrequentlyeffectivelyaddressesthehighcomplexityofhighereu-karyoticproteomesistofocusonaspecificsubproteome(targetedproteomics).Thekeyfactorinpursuingtargetedproteomicsistodefineasubsetofacompleteproteomethatcanbereproduciblyisolated.Theclosermat

5、chbetweentheproteincomplexityofmostsubproteomesandavailableanalyticalmethodsmakestargetedproteomicsaveryattrac-tiveapproachtoproteomics-basedexplorationofbiologicalprocessesanddiseases.Intermsofcellularcomplexity,subproteomescanrangefromsimplemacromolecularcom-plexestocellul

6、armachinestoorganelles.Otherstrategiesfortargetingsubproteomesaretoisolatespecificclassesofproteins,suchasphosphoproteinsortyrosine-phosphorylatedproteins,glycoproteins,oroxidativelymodifiedproteins,tonameonlyafew.Frequently,severalalternativemethodsareavailableforisolatingspe

7、cificsubproteomes.Asnotedabove,oneofthemostcriticalconsiderationsisthereproducibilityofthesubproteomeisolationmethod.Anothercriticalconsiderationisthespecificityoftheisolationmethodused;sincenomethodiscompletelyspecific,animportantissuetoconsiderishowonewilldistinguishcontamina

8、ntsfromproteinsthatbelongtothetargetedsubproteome.Anotheradvantageoftargete