| RRC ID |
38798
|
| 著者 |
Tomita M, Ohkura N, Ito M, Kato T, Royce PM, Kitajima T.
|
| タイトル |
Biosynthesis of recombinant human pro-alpha 1(III) chains in a baculovirus expression system: production of disulphide-bonded and non-disulphide-bonded species containing full-length triple helices.
|
| ジャーナル |
Biochem J
|
| Abstract |
We have investigated the expression of human procollagen III by insect cells infected with a recombinant baculovirus carrying cDNA for the pro-alpha1(III) chain of type-III collagen. A high level of expression was obtained, and a small proportion of the heterologously expressed pro-alpha1(III) chains formed normally disulphide-bonded procollagen III, which was secreted into the culture medium. This species displayed a melting temperature (Tm) of approx. 38 degrees C as assessed by its resistance to digestion by a mixture of trypsin and chymotrypsin, slightly lower than that of 39.5 degrees C for procollagen III synthesized by cultured human dermal fibroblasts, and reflected a slight degree of under-hydroxylation of prolyl residues. This is possibly a consequence of the lower incubation temperature of insect cells, or of an insufficiency of prolyl hydroxylase activity within them. A significant proportion of the expressed chains formed trimeric molecules of similar thermal stability containing an apparently full-length triple-helical region, but were not disulphide-bonded and not secreted. In addition to providing a source of recombinant human procollagen III, the system promises to be useful in the study of procollagen chain association and subsequent folding.
|
| 巻・号 |
312 ( Pt 3)(Pt 3)
|
| ページ |
847-53
|
| 公開日 |
1995-12-15
|
| DOI |
10.1042/bj3120847
|
| PMID |
8554530
|
| PMC |
PMC1136192
|
| MeSH |
Amino Acids / analysis
Animals
Ascorbic Acid / pharmacology
Baculoviridae / genetics*
Base Sequence
Cell Line
Chymotrypsin / metabolism
Disulfides / chemistry*
Drug Stability
Gene Expression*
Hot Temperature
Humans
Molecular Sequence Data
Procollagen / chemistry*
Procollagen / genetics*
Procollagen / metabolism
Protein Conformation
Recombinant Proteins / biosynthesis
Spodoptera / metabolism
Trypsin / metabolism
|
| IF |
4.097
|
| 引用数 |
30
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
Sf9(RCB0563) |
