論文 - 詳細
| RRC ID | 4923 |
|---|---|
| 著者 | Hullmann J, Patzer SI, Römer C, Hantke K, Braun V. |
| タイトル | Periplasmic chaperone FkpA is essential for imported colicin M toxicity. |
| ジャーナル | Mol Microbiol |
| Abstract |
Chaperones facilitate correct folding of newly synthesized proteins. We show here that the periplasmic FkpA chaperone is required for killing Escherichia coli by colicin M entering cells from the outside. Highly active colicin M preparations were inactive against fkpA mutant cells; 10(4)-fold dilutions killed fkpA(+) cells. Three previously isolated spontaneous mutants tolerant to colicin M carried a stop codon or an IS1 insertion in the peptidyl-prolyl-cis-trans-isomerase (PPIase) domain (C-domain) of FkpA, which resulted in deletion of the domain. A randomly generated mutant carried a G148D mutation in the C-domain. A temperature-sensitive mutant tolerant to colicin M carried a Y25N mutation in the FkpA N-domain. Mutants transformed with wild-type fkpA were colicin M-sensitive. Isolated FkpA-His reduced colicin M-His cleavage by proteinase K and renatured denatured colicin M-His in vitro; renaturation was prevented by the PPIase inhibitor FK506. In both assays, periplasmic SurA-His had no effect. No other tested periplasmic chaperone could activate colicin M. Among the tested colicins, only colicin M required FkpA for activity. Colicin M bound to cells via FhuA was inactivated by trypsin; unbound colicin M retained activity. We propose that colicin M unfolds during import across the outer membrane, FkpA specifically assists in folding colicin M into an active toxin in the periplasm and PPIase is essential for colicin M activity. Colicin M is a suitable tool for the isolation of FkpA mutants used to elucidate the functions of the FkpA N- and C-domains. |
| 巻・号 | 69(4) |
| ページ | 926-37 |
| 公開日 | 2008-8-1 |
| DOI | 10.1111/j.1365-2958.2008.06327.x |
| PII | MMI6327 |
| PMID | 18554332 |
| PMC | PMC2615193 |
| MeSH | Bacteriolysis* / genetics Colicins / metabolism* Colicins / pharmacology Endopeptidase K / chemistry Escherichia coli / drug effects Escherichia coli / enzymology* Escherichia coli / genetics Escherichia coli Proteins / genetics Escherichia coli Proteins / metabolism* Gene Deletion Hot Temperature Membrane Proteins / genetics Membrane Proteins / metabolism* Molecular Chaperones / genetics Molecular Chaperones / metabolism* Peptidylprolyl Isomerase / genetics Peptidylprolyl Isomerase / metabolism* Periplasm / enzymology* Protein Conformation Protein Denaturation Protein Transport |
| IF | 3.418 |
| 引用数 | 37 |
| WOS 分野 | MICROBIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY |
| リソース情報 | |
| 原核生物(大腸菌) | ME9062(BW25113) JW0013 JW0052 JW0157 JW0427 JW0431 JW3309 JW3326 JW3584 JW4103 |