Evaluation of non-reductive ß-elimination/Michael addition for glycosylation site determination in mucin-like O-glycopeptides.
Electrophoresis
; 32(24): 3546-53, 2011 Dec.
Article
em En
| MEDLINE
| ID: mdl-22180207
ABSTRACT
Investigation of site-specific protein O-glycosylation remains a formidable task in post-translational modification-centred proteomics. In particular, the determination of O-glycosylated amino acids in mucin-like glycopeptides lags far behind the techniques for phosphorylation site and N-glycosylation site identification, for which well-established enrichment techniques are available. The present work investigated ß-elimination of mucin-like O-glycopeptides with a mild alkylamine base and concomitant Michael-type addition using 2-mercaptoethanol as nucleophile applied to synthetic GalNAcylated O-glycopeptides as well as exoglycosidase-treated endogenous peptides isolated from human blood plasma. This strategy permits O-glycosylated sites to be unambiguously localized, even in multiple-glycosylated peptides. Peptides covalently modified with the glycan surrogate exhibit excellent backbone fragmentation in MS/MS due to their stability during CID.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Glicopeptídeos
Limite:
Humans
Idioma:
En
Revista:
Electrophoresis
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Áustria