研究人血浆中的O-连接蛋白糖基化修饰。

Studying O-linked protein glycosylations in human plasma.

作者信息

Williams Taufika Islam, Saggese Diana A, Muddiman David C

机构信息

W.M. Keck FT-ICR Mass Spectrometry Laboratory, Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA.

出版信息

J Proteome Res. 2008 Jun;7(6):2562-8. doi: 10.1021/pr800066e. Epub 2008 Apr 19.

DOI:10.1021/pr800066e

PMID:18422354

Abstract

Recent investigations have implicated aberrant glycosylations in various malignancies, including epithelial ovarian cancer (EOC). The protocol here identifies O-linked carbohydrate patterns in EOC plasma glycoproteins through chemical cleavage and purification of these glycans. Dialyzed plasma is subjected to reductive beta-elimination with alkaline borohydride to release O-linked oligosaccharides from glycoproteins. Enrichment of released glycans, as well as removal of peptide and other contaminants, is followed by carbohydrate pattern analysis with MALDI-FT-ICR-MS.

摘要

最近的研究表明，异常糖基化与包括上皮性卵巢癌（EOC）在内的各种恶性肿瘤有关。本文所述方案通过化学切割和纯化这些聚糖来鉴定EOC血浆糖蛋白中的O-连接碳水化合物模式。将透析后的血浆用碱性硼氢化物进行还原性β-消除反应，以从糖蛋白中释放出O-连接寡糖。在对释放出的聚糖进行富集以及去除肽和其他污染物之后，采用基质辅助激光解吸电离傅里叶变换离子回旋共振质谱（MALDI-FT-ICR-MS）进行碳水化合物模式分析。