Publication: Role of Glycosyltransferase 25 Domain 1 in Type i Collagen Glycosylation and Molecular Phenotypes
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Issued Date
2019
Resource Type
File Type
application/pdf
ISSN
62960
Other identifier(s)
2-s2.0-85076271790
Rights Holder(s)
มหาวิทยาลัยศรีนครินทรวิโรฒ
Bibliographic Citation
Biochemistry. Vol 58, No.50 (2019), p.5040-5051
Suggested Citation
Terajima M., Taga Y., Sricholpech M., Kayashima Y., Sumida N., Maeda N., Hattori S., Yamauchi M. Role of Glycosyltransferase 25 Domain 1 in Type i Collagen Glycosylation and Molecular Phenotypes. Biochemistry. Vol 58, No.50 (2019), p.5040-5051. doi:10.1021/acs.biochem.8b00984 Retrieved from: https://hdl.handle.net/20.500.14740/5003
Abstract
Glycosylation in type I collagen occurs as O-linked galactosyl- (G-) lesser and glucosylgalactosyl-hydroxylysine (GG-Hyl); however, its biological significance is still not well understood. To investigate the function of this modification in bone, we have generated preosteoblast MC3T3-E1 (MC)-derived clones, short hairpin (Sh) clones, in which Glt25d1 gene expression was stably suppressed. In Sh clones, the GLT25D1 protein levels were markedly diminished in comparison to controls (MC and those transfected with the empty vector). In Sh collagen, levels of both G- and GG-Hyl were significantly diminished with a concomitant increase in the level of free-Hyl. In addition, the level of immature divalent cross-links significantly diminished while the level of the mature trivalent cross-link increased. As determined by mass spectrometric analysis, seven glycosylation sites were identified in type I collagen and the most predominant site was at the helical cross-linking site, α1-87. At all of the glycosylation sites, the relative levels of G- and GG-Hyl were markedly diminished, i.e., by ∼50-75%, in Sh collagen, and at five of these sites, the level of Lys hydroxylation was significantly increased. The collagen fibrils in Sh clones were larger, and mineralization was impaired. These results indicate that GLT25D1 catalyzes galactosylation of Hyl throughout the type I collagen molecule and that this modification may regulate maturation of collagen cross-linking, fibrillogenesis, and mineralization. Copyright © 2019 American Chemical Society.
Subject(s)
Cloning
Gene expression
Glycosylation
Mass spectrometry
Mineralogy
Biological significance
Collagen fibrils
Galactosylation
Glycosyl transferase
Glycosylation sites
Mass spectrometric analysis
Molecular phenotypes
Type I collagen
Collagen
Collagen type 1
Glycosyltransferase
Short hairpin RNA
Colgalt1 protein, mouse
Collagen type 1
Galactosyltransferase
Lysine
Alpha helix
Animal cell
Article
Cell cloning
Controlled study
Gene expression
Hydroxylation
In vitro study
Mass spectrometry
MC3T3-E1 cell line
Molecular cloning
Mouse
Nonhuman
Priority journal
Protein analysis
Protein cross linking
Protein domain
Protein glycosylation
3T3 cell line
Animal
Biocatalysis
Chemistry
Glycosylation
Metabolism
Phenotype
3T3 Cells
Animals
Biocatalysis
Collagen Type I
Galactosyltransferases
Glycosylation
Lysine
Mice
Phenotype
Gene expression
Glycosylation
Mass spectrometry
Mineralogy
Biological significance
Collagen fibrils
Galactosylation
Glycosyl transferase
Glycosylation sites
Mass spectrometric analysis
Molecular phenotypes
Type I collagen
Collagen
Collagen type 1
Glycosyltransferase
Short hairpin RNA
Colgalt1 protein, mouse
Collagen type 1
Galactosyltransferase
Lysine
Alpha helix
Animal cell
Article
Cell cloning
Controlled study
Gene expression
Hydroxylation
In vitro study
Mass spectrometry
MC3T3-E1 cell line
Molecular cloning
Mouse
Nonhuman
Priority journal
Protein analysis
Protein cross linking
Protein domain
Protein glycosylation
3T3 cell line
Animal
Biocatalysis
Chemistry
Glycosylation
Metabolism
Phenotype
3T3 Cells
Animals
Biocatalysis
Collagen Type I
Galactosyltransferases
Glycosylation
Lysine
Mice
Phenotype