采用质谱法进行全甲基化N-聚糖分析。

Permethylated N-glycan analysis with mass spectrometry.

作者信息

Lin Zhenxin, Lubman David M

机构信息

Department of Chemistry, University of Michigan, Ann Arbor, MI, USA.

出版信息

Methods Mol Biol. 2013;1007:289-300. doi: 10.1007/978-1-62703-392-3_12.

DOI:10.1007/978-1-62703-392-3_12

PMID:23666731

Abstract

Protein glycosylation plays an important role in multiple cell functions, and aberrations of protein glycosylation are associated with various malignancies including cancer. In this chapter, we provide a detailed protocol for MALDI MS analysis of permethylated N-glycans extracted from human serum proteins. The protocol includes procedures for N-glycan purification and in-solution permethylation, structural elucidation of permethylated N-glycans by MALDI-QIT-TOF MS, and construction of indices to quantify levels of certain types of glycosylation, such as fucosylation, which may serve as a potential disease biomarker.

摘要

蛋白质糖基化在多种细胞功能中发挥着重要作用，蛋白质糖基化异常与包括癌症在内的各种恶性肿瘤相关。在本章中，我们提供了一份详细的方案，用于对从人血清蛋白中提取的全甲基化N-聚糖进行基质辅助激光解吸电离质谱（MALDI MS）分析。该方案包括N-聚糖纯化和溶液内全甲基化的步骤、通过MALDI-四极杆离子阱-飞行时间质谱（MALDI-QIT-TOF MS）对全甲基化N-聚糖进行结构解析，以及构建指数以量化某些类型糖基化（如岩藻糖基化）的水平，岩藻糖基化可能作为一种潜在的疾病生物标志物。