Development of a method to analyze glycosylation site-specific glycome at proteome-scale for study on significance of glycan heterogeneity

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K14426
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Genome biology
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: Kaji Hiroyuki 国立研究開発法人産業技術総合研究所, 創薬基盤研究部門, 研究グループ長 (80214302)
• Research Collaborator: TOMIOKA Azusa ; FUJITA Mika ; SUKEGAWA Masako ; OKATANI Chiaki ; SHIKANAI Toshihide ; NARIMATSU Hisashi
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: グライコプロテオミクス / 糖鎖修飾 / 翻訳後修飾 / 質量分析 / 糖鎖不均一性 / マウス / グライコーム / プロテオミクス

Outline of Final Research Achievements

Since glycans on glycoproteins are synthesized by many glycan synthesis-related enzymes, they have diverse and heterogeneous structures. Since the expression profile of the enzyme varies depending on the cell type, the glycan variety (glycomes) varies from cell to cell. Furthermore, glycomes are recognized to be different for each type of protein, and at each attachment site. However, little is known about the actual state of glycan heterogeneity. Direct analysis of glycopeptides is essential for analyzing site-specific glycomes, and several methods using MS analysis based on multi-step dissociation have been developed. However, its sensitivity is quite low. Therefore, we developed an MS/MS-independent method for site-specific glycome analysis, and applied it to the glycopeptides obtained from several major mouse tissues. We could obtain large scale information of site specific glycomes; about 12,000 site-specific glycoforms at about 1,200 sites on ca 800 glycoproteins.

Free Research Field

プロテオミクス