Minuth, Torsten and Henn, Martina and Rutkat, Kerstin and Andrae, Stefan and Frey, Gerhard and Rachel, Reinhard and Stetter, Karl Otto and Jaenicke, Rainer (1999) The recombinant thermosome from the hyperthermophilic archaeon Methanopyros kandleri: In vitro analysis of its chaperone activity. BIOLOGICAL CHEMISTRY, 380 (1). pp. 55-62. ISSN 1431-6730,
Full text not available from this repository.Abstract
The archaeon Methanopyrus kandleri is the most thermophilic methanogen presently known. It contains a chaperonin (thermosome) which represents a 951 kDa homo-hexadecameric protein complex with NH4+-dependent ATPase activity, Since its synthesis is not increased upon heat shock, we set out to test its chaperone function. In order to obtain the chaperonin in amounts sufficient for functional investigations, the gene encoding the 60 kDa subunit was expressed in E. coli BL21(DE3) cells. Purification yielded soluble, high-molecular-mass double-ring complexes, indistinguishable from the natural thermosome. in order to study the functional properties of the recombinant protein complex, pig citrate synthase, yeast alcohol dehydrogenase, yeast alpha-glucosidase, bovine insulin, and Thermotoga phosphoglycerate kinase were used as model substrates. The results demonstrate that the recombinant M. kandleri thermosome possesses a chaperone-like activity in vitro, inhibiting aggregation as the major off-pathway-reaction during thermal unfolding and refolding of proteins after chemical denaturation. However, the chaperonin only forms dead-end complexes with its non-native substrates, no release is detectable at temperatures between 25 and 60 degrees C.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | GROUP-II CHAPERONIN; IN-VITRO; HEAT-SHOCK; TETRAHYDROMETHANOPTERIN FORMYLTRANSFERASE; MOLECULAR CHAPERONE; SALT-DEPENDENCE; PROTEIN; AGGREGATION; BINDING; FORMYLMETHANOFURAN; chaperonin; hyperthermophilic methanogen; Methanopyrus kandleri; protein aggregation; thermosome |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) > Prof. Dr. Reinhard Rachel |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 29 Nov 2022 10:03 |
| Last Modified: | 29 Nov 2022 10:03 |
| URI: | https://pred.uni-regensburg.de/id/eprint/48615 |
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