Analysis of Glycopeptides Using Lectin Affinity Chromatography with MALDI-TOF Mass Spectrometry

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• 作者：Kazutosi Kubota ; Yuji Sato ; Yusuke Suzuki ; Naoko Goto-Inoue ; Tosifusa Toda ; Minoru Suzuki ; Shin-ichi Hisanaga ; Akemi Suzuki ; Tamao Endo
• 刊名：Analytical Chemistry
• 出版年：2008
• 出版时间：May 15, 2008
• 年：2008
• 卷：80
• 期：10
• 页码：3693 - 3698
• 全文大小：385K
• 年卷期：v.80,no.10(May 15, 2008)
• ISSN：1520-6882

文摘

Glycopeptides prepared from 1 nmol of a mixture of glycoproteins, transferrin, and ribonuclease B by lysylendopeptidase 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 MSⁿ 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.