title: Glycosylation and cross-linking in bone type I collagen.
abstract: Fibrillar type I collagen is the major organic component in bone, providing a stable template for mineralization. During collagen biosynthesis, specific hydroxylysine residues become glycosylated in the form of galactosyl- and glucosylgalactosyl-hydroxylysine. Furthermore, key glycosylated hydroxylysine residues, α1/2-87, are involved in covalent intermolecular cross-linking. Although cross-linking is crucial for the stability and mineralization of collagen, the biological function of glycosylation in cross-linking is not well understood. In this study, we quantitatively characterized glycosylation of non-cross-linked and cross-linked peptides by biochemical and nanoscale liquid chromatography-high resolution tandem mass spectrometric analyses. The results showed that glycosylation of non-cross-linked hydroxylysine is different from that involved in cross-linking. Among the cross-linked species involving α1/2-87, divalent cross-links were glycosylated with both mono- and disaccharides, whereas the mature, trivalent cross-links were primarily monoglycosylated. Markedly diminished diglycosylation in trivalent cross-links at this locus was also confirmed in type II collagen. The data, together with our recent report (Sricholpech, M., Perdivara, I., Yokoyama, M., Nagaoka, H., Terajima, M., Tomer, K. B., and Yamauchi, M. (2012) Lysyl hydroxylase 3-mediated glucosylation in type I collagen: molecular loci and biological significance. J. Biol. Chem. 287, 22998-23009), indicate that the extent and pattern of glycosylation may regulate cross-link maturation in fibrillar collagen.
authors: North Carolina Oral Health Institute, School of Dentistry, University of North Carolina, Chapel Hill, North Carolina 27599.
authors: Masahiko
authors: M
authors: Terajima
authors: Laboratory of Structural Biology, NIEHS, National Institutes of Health, Research Triangle Park, North Carolina 27709.
authors: Irina
authors: I
authors: Perdivara
authors: Department of Oral Surgery and Oral Medicine, Faculty of Dentistry, Srinakharinwirot University, Bangkok 10110, Thailand, and.
authors: Marnisa
authors: M
authors: Sricholpech
authors: North Carolina Oral Health Institute, School of Dentistry, University of North Carolina, Chapel Hill, North Carolina 27599.
authors: Yoshizumi
authors: Y
authors: Deguchi
authors: Tissue Imaging and Spectroscopy Laboratory, Department of Bioengineering, Temple University, Philadelphia, Pennsylvania 19122.
authors: Nancy
authors: N
authors: Pleshko
authors: Laboratory of Structural Biology, NIEHS, National Institutes of Health, Research Triangle Park, North Carolina 27709.
authors: Kenneth B
authors: KB
authors: Tomer
authors: North Carolina Oral Health Institute, School of Dentistry, University of North Carolina, Chapel Hill, North Carolina 27599,. Electronic address: yamauchm@dentistry.unc.edu.
authors: Mitsuo
authors: M
authors: Yamauchi
mesh_terms: D000818:Animals; D001842:Bone and Bones / Q000737:chemistry*; D002417:Cattle; D002853:Chromatography, Liquid; D024042:Collagen Type I / Q000737:chemistry*; D006031:Glycosylation; D006901:Hydroxylysine / Q000737:chemistry*; D013058:Mass Spectrometry; D055550:Protein Stability
chemical_list: D024042:Collagen Type I; D006901:Hydroxylysine
keywords: Bone; Collagen; Glycosylation; Hydroxylysine (Hyl); Mass Spectrometry (MS); Post-translational Modification (PTM)