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 language | English |
|---|---|
| Pages (from-to) | 187-196 |
| Number of pages | 10 |
| Journal | Cell Structure and Function |
| Volume | 24 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - 1999 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Effect of HSP47 on prolyl 4-hydroxylation of collagen model peptides
AU - Asada, Shinichi
AU - Koide, Takaki
AU - Yasui, Hiroyuki
AU - Nagata, Kazuhiro
PY - 1999
Y1 - 1999
N2 - 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.
AB - 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.
KW - 4-Hydroxy-L-proline
KW - Collagen
KW - HSP47
KW - Peptide
KW - Prolyl 4-hydroxylase
UR - http://www.scopus.com/inward/record.url?scp=0032751783&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032751783&partnerID=8YFLogxK
U2 - 10.1247/csf.24.187
DO - 10.1247/csf.24.187
M3 - Article
C2 - 10532353
AN - SCOPUS:0032751783
VL - 24
SP - 187
EP - 196
JO - Cell Structure and Function
JF - Cell Structure and Function
SN - 0386-7196
IS - 4
ER -