Analysis of glycopeptides using lectin affinity chromatography with MALDI-TOF mass spectrometry

被引：48

作者：

Kubota, Kazutosi ^{[1
,4
]}

Sato, Yuji ^{[1
]}

Suzuki, Yusuke ^{[3
]}

Goto-Inoue, Naoko ^{[3
]}

Toda, Tosifusa ^{[2
]}

Suzuki, Minoru ^{[3
]}

Hisanaga, Shin-ichi ^{[4
]}

Suzuki, Akemi ^{[3
]}

Endo, Tamao ^{[1
]}

机构：

[1] Tokyo Metropolitan Inst Gerontol, Res Team Funct Genom, Fdn Res Aging & Promot Human Welf, Itabashi Ku, Tokyo 1730015, Japan

[2] Tokyo Metropolitan Inst Gerontol, Res Team Mol Biomarkers, Fdn Res Aging & Promot Human Welf, Itabashi Ku, Tokyo 1730015, Japan

[3] RIKEN, Sphingolipid Express Lab, Inst Phys & Chem Res, Wako, Saitama 3510198, Japan

[4] Tokyo Metropolitan Univ, Dept Biol Sci, Grad Sch Sci, Tokyo 1920397, Japan

来源：

ANALYTICAL CHEMISTRY | 2008年 / 80卷 / 10期

关键词：

D O I：

10.1021/ac800070d

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Glycopeptides prepared from 1 nmol of a mixture of glycoproteins, transferrin, and ribonuclease B by lysylen-dopeptidase digestion were isolated by lectin and cellulose column chromatographies, and then they were analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and MALDI-quadrupole ion trap (QIT)-TOF mass spectrometry which enables the performance of MS' analysis. The lectin affinity preparation of glycopeptides with Sambucus nigra agglutinin and concanavalin A provides the glycan structure outlines for the sialyl linkage and the core structure of N-glycans. Such structural estimation was confirmed by MALDI-TOF MS and MALDI-QIT-TOF MS/MS. Amino acid sequences and location of glycosylation sites were determined by MALDI-QIT-TOF MS/MS/MS. Taken together, the combination of lectin column chromatography, MALDI-TOF MS, and MALDI-QIT-TOF MSn provides an easy way for the structural estimation of glycans and the rapid analysis of glycoproteomics.

引用

页码：3693 / 3698

页数：6

共 36 条

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