| RRC ID |
30171
|
| 著者 |
Miyazaki T, Yoshimi T, Furutsu Y, Hongo K, Mizobata T, Kanemori M, Kawata Y.
|
| タイトル |
GroEL-substrate-GroES ternary complexes are an important transient intermediate of the chaperonin cycle.
|
| ジャーナル |
J Biol Chem
|
| Abstract |
GroEL C138W is a mutant form of Escherichia coli GroEL, which forms an arrested ternary complex composed of GroEL, the co-chaperonin GroES and the refolding protein molecule rhodanese at 25 degrees C. This state of arrest could be reversed with a simple increase in temperature. In this study, we found that GroEL C138W formed both stable trans- and cis-ternary complexes with a number of refolding proteins in addition to bovine rhodanese. These complexes could be reactivated by a temperature shift to obtain active refolded protein. The simultaneous binding of GroES and substrate to the cis ring suggested that an efficient transfer of substrate protein into the GroEL central cavity was assured by the binding of GroES prior to complete substrate release from the apical domain. Stopped-flow fluorescence spectroscopy of the mutant chaperonin revealed a temperature-dependent conformational change in GroEL C138W that acts as a trigger for complete protein release. The behavior of GroEL C138W was reflected closely in its in vivo characteristics, demonstrating the importance of this conformational change to the overall activity of GroEL.
|
| 巻・号 |
277(52)
|
| ページ |
50621-8
|
| 公開日 |
2002-12-27
|
| DOI |
10.1074/jbc.M209183200
|
| PII |
S0021-9258(19)31378-X
|
| PMID |
12377767
|
| MeSH |
Amino Acid Substitution
Animals
Chaperonin 10 / genetics
Chaperonin 10 / metabolism*
Chaperonin 60 / genetics
Chaperonin 60 / metabolism*
Chaperonins / metabolism*
Enzymes / metabolism
Escherichia coli Proteins / genetics
Escherichia coli Proteins / metabolism*
Kinetics
Macromolecular Substances
Operon
Protein Folding
Recombinant Proteins / metabolism
Thiosulfate Sulfurtransferase / chemistry
Thiosulfate Sulfurtransferase / metabolism*
alpha-Amylases / metabolism
|
| IF |
4.238
|
| 引用数 |
20
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| 原核生物(大腸菌) |
ME7966 |
