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

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Title: High Throughput Quantitative Glycomics and Glycoform-focused Proteomics of Murine Dermis and Epidermis
Authors: Uematsu, Rie Browse this author
Furukawa, Jun-ichi Browse this author
Nakagawa, Hiroaki Browse this author
Shinohara, Yasuro4 Browse this author
Deguchi, Kisaburo Browse this author
Monde, Kenji Browse this author →KAKEN DB
Nishimura, Shin-Ichiro Browse this author →KAKEN DB
Authors(alt): 篠原, 康郎4
Keywords: tandem mass-spectrometry
Issue Date: 16-Sep-2005
Publisher: The American Society for Biochemistry and Molecular Biology, Inc.
Journal Title: Molecular & Cellular Proteomics
Volume: 4
Issue: 12
Start Page: 1977
End Page: 1989
Publisher DOI: 10.1074/mcp.M500203-MCP200
PMID: 16170054
Abstract: 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
Type: article (author version)
Appears in Collections:理学院・理学研究院 (Graduate School of Science / Faculty of Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 篠原 康郎

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