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Mass Spectrometry Based Glycoproteomics—From a Proteomics Perspective.full

Review

©2011byTheAmericanSocietyforBiochemistryandMolecularBiology,Inc.Thispaperisavailableonlineathttp://doc.docsou.com

MassSpectrometryBasedGlycoproteomics—FromaProteomicsPerspective*

ShengPan ,RuChen§,RuediAebersold¶,andTeresaA.Brentnall§

Glycosylationisoneofthemostimportantandcommonformsofproteinpost-translationalmodificationthatisinvolvedinmanyphysiologicalfunctionsandbiologicalpathways.Alteredglycosylationhasbeenassociatedwithavarietyofdiseases,includingcancer,inflammatoryanddegenerativediseases.Glycoproteinsarebecomingim-portanttargetsforthedevelopmentofbiomarkersfordiseasediagnosis,prognosis,andtherapeuticresponsetodrugs.Theemergingtechnologyofglycoproteomics,whichfocusesonglycoproteomeanalysis,isincreasinglybecominganimportanttoolforbiomarkerdiscovery.Anin-depth,comprehensiveidentificationofaberrantglyco-proteins,andfurther,quantitativedetectionofspecificglycosylationabnormalitiesinacomplexenvironmentre-quireaconcertedapproachdrawingfromavarietyoftechniques.Thisreportprovidesanoverviewoftherecentadvancesinmassspectrometrybasedglycoproteomicmethodsandtechnology,inthecontextofbiomarkerdis-coveryandclinicalapplication.Molecular&CellularProteomics10:10.1074/mcp.R110.003251,1–14,2011.

tocancer(4–6).Recentreportshaveprovidedacomprehen-siveoverviewoftheconceptofglycomicsanditsprospectiveinbiomarkerresearch(7–10).Incontrast,thisreviewisfo-cusedonrecentdevelopmentsinglycoproteomictechniquesandtheiruniqueapplicationandtechnicalchallengetobio-markerdiscovery.

GlycoproteomicsinBiomarkerDiscoveryandClinicalStudy—Mostsecretoryandmembrane-boundproteinspro-ducedbymammaliancellscontaincovalentlylinkedglycanswithdiversestructures(2).Theglycosylationformofaglyco-proteinishighlyspecificateachglycosylationsiteandgen-erallystableforagivencelltypeandphysiologicalstate.However,theglycosylationformofaproteincanbealteredsignificantlybecauseofchangesincellularpathwaysandprocessesresultingfromdiseases,suchascancer,inflamma-tion,andneurodegeneration.Suchdisease-associatedalter-ationsinglycoproteinscanhappeninoneorbothoftwoways:1)proteinglycosylationsitesareeitherhypo,hyper,ornewlyglycosylatedand/or;2)theglycosylationformoftheattachedcarbohydratemoietyisaltered.Infact,alteredgly-cosylationpatternshavelongbeenrecognizedashallmarksincancerprogression,inwhichtumor-specificglycoproteinsareactivelyinvolvedinneoplasticprogressionandmetastasis(5,6,11,12).Sensitivedetectionofsuchdisease-associatedglycosylationchangesandabnormalitiescanprovideauniqueavenuetodevelopglycoproteinbiomarkersfordiag-nosisandprognosis.Inaddition,interventionintheglycosyl-ationandcarbohydrate-dependentcellularpathwaysrepre-sentapotentialnewmodalityforcancertherapies(6,11,13).TableIlistssomeoftheFDAapprovedcancerbiomarkers(14,15)thatareglycosylatedproteinsorproteincomplexes.

Proteinbiomarkerdevelopmentisacomplexandchalleng-ingtask.Thecriteriaandapproachappliedfordevelopingeachindividualbiomarkercanvary,dependingonthepur-poseofthebiomarkerandtheperformancerequirementforitsclinicalapplication(16,17).Ingeneral,ithasbeensug-gestedthatthepreclinicalexploratoryphaseofproteinbio-markerdevelopmentcanbetechnicallydefinedintofourstages(18),includinginitialdiscoveryofdifferentialproteins;testingandselectionofqualifiedcandidates;verificationofasubsetofcandidates;assaydevelopmentandpre-clinicalvalidationofpotentialbiomarkers.Thankstorecenttechno-logicaladvances,massspectrometrybasedglycoproteomicsisnowplayingamajorroleintheinitialphaseofdiscovering

Withrecentadvancesinproteomics,analyticalandcom-putationaltechnologies,glycoproteomics—theglobalanaly-sisofglycoproteins—israpidlyemergingasasubfieldofproteomicswithhighbiologicalandclinicalrelevance.Glyco-proteomicsintegratesglycoproteinenrichmentandproteo-micstechnologiestosupportthesystematicidentificationandquantificationofglycoproteinsinacomplexsample.There-centdevelopmentofthesetechniqueshasstimulatedgreatinterestinapplyingthetechnologyinclinicaltranslationalstudies,inparticular,proteinbiomarkerresearch.

Whileglycomicsisthestudyofglycome(repertoireofgly-cans),glycoproteomicsfocusesonstudyingtheprofileofglycosylatedproteins,i.e.theglycoproteome,inabiologicalsystem.Considerableworkhasbeendonetocharacterizethesequencesandprimarystructureoftheglycanmoietiesat-tachedtoproteins(1–3),andtheirstructuralalterationsrelated

Fromthe DepartmentofPathology,UniversityofWashington,Seattle,WA98195,§DivisionofGastroenterology/DepartmentofMedicine,UniversityofWashington,Seattle,WA98195,¶InstituteofMolecularSystemsBiology,ETHZurichandFacultyofScience,UniversityofZurich,Switzerland

ReceivedAugust27,2010,andinrevisedform,September2,2010Published,MCPPapersinPress,August27,2010,DOI10.1074/mcp.R110.003251

Molecular&CellularProteomics10.110.1074/mcp.R110.003251–1

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