A functional role of the glycosylated N-terminal domain of chondromodulin-I.
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概要
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Chondromodulin-I (ChM-I) is a 25-kDa glycoprotein that specifically localizes in the extracellular matrix of cartilage and negatively regulates angiogenesis. ChM-I comprises two domains: an N-terminal hydrophilic domain (domain 1) containing an N-linked glycosylation site and a C-terminal hydrophobic domain (domain 2) with all four disulfide bonds that are present in this protein. We generated a nonglycosylated recombinant human ChM-I (NG-hChM-I) and compared its bioactivity with that of the glycosylated form of human ChM-I (G-hChM-I) expressed in Chinese hamster ovary cells in vitro. NG-hChM-I exhibited the growth factor/inhibitor activity in the cultures of chondrocytes and vascular endothelial cells but required markedly higher doses. Although domain 1 is predicted to be hydrophilic per se on the basis of its amino acid sequence, NG-hChM-I remains insoluble in aqueous solution as much as ΔN-hChM-I that lacks the N-terminal 37 amino acids containing an N-glycosylation site. Circular dichroism measurements revealed that the content of α-helix was calculated to be 34% in G-hChM-I, whereas the content of the characteristic secondary structures in NG-hChM-I was distinctly lower than those in G-hChM-I. These results indicate that glycosylation in domain 1 is critical for the structural integrity for biological functions of ChM-I in vitro.
- 2011-01-30
著者
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Hiraki Yuji
Department of Cellular Differentiation, Institute for Frontier Medical Sciences, Kyoto University
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Sato Koji
Department of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, T
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Kondo J
Research Center Mitsubishi Pharma Corporation
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Hiraki Y
慶応義塾大学 循環器内科学
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Hiraki Yuji
Department Of Biochemistry Osaka University Faculty Of Dentistry
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Shibata Hiroyuki
Department Of Cardiology Himeji Cardiovascular Center
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Shukunami Chisa
Department Of Molecular Interaction And Tissue Engineering Institute For Frontier Medical Sciences K
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Shukunami Chisa
Department Of Cellular Differentiation Institute For Frontier Medical Sciences Kyoto University
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Miura Shigenori
Department Of Cellular Differentiation Institute For Frontier Medical Sciences Kyoto University
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Kondo Jun
Research Center Mitsubishi Pharma Corporation
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Higuchi Yoshiki
Department Of Life Science Graduate School Of Life Science University Of Hyogo
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KONDO Jun
Advanced Medical Research Laboratory, Research Division, Mitsubishi Tanabe Pharma Corporation
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YAMAKAWA Akira
Department of Life Science, Graduate School of Life Science, University of Hyogo
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Yamakawa Akira
Department Of Life Science Graduate School Of Life Science University Of Hyogo
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Sato Koji
Department Of Cellular Differentiation Institute For Frontier Medical Sciences Kyoto University
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Sato Koji
Department Of Bioenvironmental Medicine Graduate School Of Medicine Chiba University
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Shibata Hiroyuki
Department Of Cellular Differentiation Institute For Frontier Medical Sciences Kyoto University
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Sato Koji
Department Of Anesthesia And Intensive Care Medicine Akita University Graduate School Of Medicine
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HIRAKI Yuji
Department of Biochemistry and Calcified Tissue Metabolism, Faculty of Dentistry
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