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High Throughput Quantitative Glycomics and Glycoform-focused Proteomics of Murine Dermis and Epidermis

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タイトル: High Throughput Quantitative Glycomics and Glycoform-focused Proteomics of Murine Dermis and Epidermis
著者: Uematsu, Rie 著作を一覧する
Furukawa, Jun-ichi 著作を一覧する
Nakagawa, Hiroaki 著作を一覧する
Shinohara, Yasuro 著作を一覧する
Deguchi, Kisaburo 著作を一覧する
Monde, Kenji 著作を一覧する
Nishimura, Shin-Ichiro 著作を一覧する
キーワード: tandem mass-spectrometry
n-linked
glycoroteins
electron-capture
lectin
acid
skin
oligosacchrides
differentiation
idendification
発行日: 2005年 9月16日
出版者: The American Society for Biochemistry and Molecular Biology, Inc.
誌名: Molecular & Cellular Proteomics
巻: 4
号: 12
開始ページ: 1977
終了ページ: 1989
出版社 DOI: 10.1074/mcp.M500203-MCP200
抄録: Despite recent advances in our understanding of the significance of the protein glycosylation, the throughput of protein glycosylation analysis is still too low to be applied to the exhaustive glycoproteomic analysis. Aiming to elucidate the N-glycosylation of murine epidermis and dermis glycoproteins, here we used a novel approach for focused proteomics. A gross N-glycan profiling (glycomics) of epidermis and dermis was first elucidated both qualitatively and quantitatively upon N-glycan derivatization with novel, stable isotope-coded derivatization reagents followed by MALDI-TOF(/TOF) analysis. This analysis revealed distinct features of the N-glycosylation profile of epidermis and dermis for the first time. A high abundance of high mannose type oligosaccharides was found to be characteristic of murine epidermis glycoproteins. Based on this observation, we performed high mannose type glycoform-focused proteomics by direct tryptic digestion of protein mixtures and affinity enrichment. We identified 15 glycoproteins with 19 N-glycosylation sites that carry high mannose type glycans by off-line LC-MALDI-TOF/TOF mass spectrometry. Moreover the relative quantity of microheterogeneity of different glycoforms present at each N-glycan binding site was determined. Glycoproteins identified were often contained in lysosomes (e.g. cathepsin L and gamma-glutamyl hydrolase), lamellar granules (e.g. glucosylceramidase and cathepsin D), and desmosomes (e g. desmocollin 1, desmocollin 3, and desmoglein). Lamellar granules are organelles found in the terminally differentiating cells of keratinizing epithelia, and desmosomes are intercellular junctions in vertebrate epithelial cells, thus indicating that N-glycosylation of tissue-specific glycoproteins may contribute to increase the relative proportion of high mannose glycans. The striking roles of lysosomal enzymes in epidermis during lipid remodeling and desquamation may also reflect the observed high abundance of high mannose glycans.
Rights: Copyright © 2005 by the American Society for Biochemistry and Molecular Biology
資料タイプ: article (author version)
URI: http://hdl.handle.net/2115/899
出現コレクション:雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

提供者: 篠原 康郎

 

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