基于超滤膜辅助的糖蛋白全N-连接糖链的富集和质谱解析(英文)

基于超滤膜辅助的糖蛋白全N-连接糖链的富集和质谱解析(英文)Based on the Enrichment and Mass Spectrometry Analysis of the Entire N

N- 基于超滤膜辅助的糖蛋白全连接糖链的富集和质 () 谱解析英文 BasedontheEnrichmentandMassSpectrometryAnalysisof theEntireN-LinkedGlycanChainsofGlycoproteinsAssistedby UltrafiltrationMembrane Introduction: Glycoproteinsareagroupofproteinsthathave oligosaccharidechainscovalentlyattachedtotheirprotein backbone.N-glycosylationisoneofthemostcommon glycosylationtypesineukaryoticproteins,whichinvolvesthe transferofglycanchainsontotheamidenitrogenofasparagine residueswithintheAsn-X-Ser/Thrsequonmotif.TheN-glycan chainsareinvolvedinawiderangeofbiologicalprocesses, includingcellrecognition,signaltransduction,immuneresponse, andproteinfoldingandstability.Therefore,thecharacterizationof theN-glycanstructuresofglycoproteinsisofgreatsignificance forunderstandingtheirfunctionanddesigningtherapeuticdrugs. However,theanalysisofN-glycanstructuresisstill challengingduetothestructuraldiversityandheterogeneityof glycoproteinsandtheirN-glycans.Typically,theN-glycansare releasedfromtheproteinbackbonebyenzymaticcleavage, followedbyderivatizationandchromatographicseparationbefore massspectrometryanalysis.However,thisapproachusuallyresults inthelossofinformationabouttheintactN-glycanstructures, especiallyforlargeglycoproteinswithcomplexN-glycanpatterns. Therefore,alternativemethodsforthecomprehensiveanalysisof N-glycanstructuresofglycoproteinsareneeded. Ultrafiltrationmembrane-basedmethodshaveemergedas promisingtoolsforglycananalysisinrecentyears.The ultrafiltrationmembraneshavedefinedporesizesthatcan separatemacromoleculesbasedontheirmolecularweight, allowingfortheselectiveenrichmentofglycanchainsfromthe proteinmixture.Inthisstudy,wedevelopedanultrafiltration membrane-basedmethodfortheenrichmentandmass spectrometryanalysisoftheentireN-linkedglycanchainsof glycoproteins.