Effect of HSP47 on prolyl 4-hydroxylation of collagen model peptides

Shinichi Asada, Takaki Koide, Hiroyuki Yasui, Kazuhiro Nagata

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Prolyl 4-hydroxylation, the most important post-translational modification in collagen biosynthesis, is catalyzed by prolyl 4-hydroxylase, an endoplasmic reticulum-resident enzyme. HSP47 is a collagen-binding stress protein which also resides in the endoplasmic reticulum. Both prolyl 4-hydroxylase and HSP47 interact with procollagen α-chains during their folding and/or modification in the endoplasmic reticulum. Recent study has revealed that a simple collagen model peptide, (Pro-Pro-Gly)(n), is recognized by HSP47 as well as by prolyl 4-hydroxylase in vitro. In the present study, we investigated the effect of HSP47 on the prolyl 4-hydroxylation of such collagen model peptides. To monitor the enzymatic hydroxylation of the peptides, we developed a non-RI assay system based on reversed-phase HPLC. When HSP47 was added to the reaction mixture, substrate and less-hydroxylated materials accumulated. This effect depended on the peptide-binding activity of HSP47, because a mutant HSP47 without collagen-binding activity did not show any inhibitory effect on prolyl 4-hydroxylation. Kinetic analysis and other biochemical analyses suggest that HSP47 retards the enzymatic reaction competing for the substrate peptide.

Original languageEnglish
Pages (from-to)187-196
Number of pages10
JournalCell Structure and Function
Volume24
Issue number4
DOIs
Publication statusPublished - 1999
Externally publishedYes

Fingerprint

Hydroxylation
Prolyl Hydroxylases
Collagen
Peptides
Endoplasmic Reticulum
prolyl-prolyl-glycine
Procollagen
Post Translational Protein Processing
Heat-Shock Proteins
Carrier Proteins
High Pressure Liquid Chromatography
Enzymes

Keywords

  • 4-Hydroxy-L-proline
  • Collagen
  • HSP47
  • Peptide
  • Prolyl 4-hydroxylase

ASJC Scopus subject areas

  • Physiology
  • Structural Biology
  • Cell Biology

Cite this

Effect of HSP47 on prolyl 4-hydroxylation of collagen model peptides. / Asada, Shinichi; Koide, Takaki; Yasui, Hiroyuki; Nagata, Kazuhiro.

In: Cell Structure and Function, Vol. 24, No. 4, 1999, p. 187-196.

Research output: Contribution to journalArticle

Asada, Shinichi ; Koide, Takaki ; Yasui, Hiroyuki ; Nagata, Kazuhiro. / Effect of HSP47 on prolyl 4-hydroxylation of collagen model peptides. In: Cell Structure and Function. 1999 ; Vol. 24, No. 4. pp. 187-196.
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