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- Title
A functional role of the glycosylated N-terminal domain of chondromodulin-I.
- Authors
Kondo, Jun; Shibata, Hiroyuki; Miura, Shigenori; Yamakawa, Akira; Sato, Koji; Higuchi, Yoshiki; Shukunami, Chisa; Hiraki, Yuji
- Abstract
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.
- Subjects
METABOLISM; AMINO acids; ANIMAL experimentation; CARTILAGE cells; CELL culture; CELL lines; COMPARATIVE studies; DOCUMENTATION; EPITHELIAL cells; ESCHERICHIA coli; GLYCOSYLATION; GROWTH factors; HAMSTERS; RESEARCH methodology; MEDICAL cooperation; MEMBRANE proteins; MICE; NEOVASCULARIZATION inhibitors; RESEARCH; RODENTS; SPECTRUM analysis; EVALUATION research; PHARMACODYNAMICS
- Publication
Journal of Bone & Mineral Metabolism, 2011, Vol 29, Issue 1, p23
- ISSN
0914-8779
- Publication type
journal article
- DOI
10.1007/s00774-010-0193-0