Characterization of cellobiohydrolase I N-glycans and differentiation of their phosphorylated isomers by capillary electrophoresis-Q-Trap mass spectrometry.
Anal Chem
; 76(19): 5878-86, 2004 Oct 01.
Article
em En
| MEDLINE
| ID: mdl-15456310
ABSTRACT
A capillary electrophoresis-mass spectrometric (CE-MS) method is described for the simultaneous analysis of uncharged and charged glycans. The glycans were labeled with the negatively charged tag 8-aminopyrene-1,3,6-trisulfonate by reductive amination and separated in an ammonium acetate buffer. A Q-Trap instrument was used for mass spectrometric detection. The CE-MS method was first optimized using maltooligosaccharides and ribonuclease B N-glycans and then applied to the characterization of enzymatically released N-glycans from the glycoprotein cellobiohydrolase I. The method, as developed, allowed differentiation of phosphorylated isomers and MS/MS provided useful structural information. Further structural evidence was obtained by studying the methylated glycans in off-line ESI-MS/MS experiments and by using a combination of chemical and enzymatic sequencing.
Buscar no Google
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Polissacarídeos
/
Espectrometria de Massas
/
Eletroforese Capilar
/
Celulose 1,4-beta-Celobiosidase
Idioma:
En
Revista:
Anal Chem
Ano de publicação:
2004
Tipo de documento:
Article