Lingl, A and Huber, H and Stetter, KO and Mayer, F and Kellermann, J and Muller, V (2003) Isolation of a complete A(1)A(0) ATP synthase comprising nine subunits from the hyperthermophile Methanococcus jannaschii. EXTREMOPHILES, 7 (3). pp. 249-257. ISSN 1431-0651,
Full text not available from this repository. (Request a copy)Abstract
Archaeal A(1)A(O) ATP synthase/ATPase operons are highly conserved among species and comprise at least nine genes encoding structural proteins. However, all A(1)A(O) ATPase preparations reported to date contained only three to six subunits and, therefore, the study of this unique class of secondary energy converters is still in its infancy. To improve the quality of A(1)A(O) ATPase preparations, we chose the hyperthermophilic, methanogenic archaeon Methanococcus jannaschii as a model organism. Individual subunits of the A(1)A(O) ATPase from M. jannaschii were produced in E. coli, purified, and antibodies were raised. The antibodies enabled the development of a protocol ensuring purification of the entire nine-subunit A(1)A(O) ATPase. The ATPase was solubilized from membranes of M. jannaschii by Triton X-100 and purified to apparent homogeneity by sucrose density gradient centrifugation, ion exchange chromatography, and gel filtration. Electron micrographs revealed the A(1) and A(O) domains and the central stalk, but also additional masses which could represent a second stalk. Inhibitor studies were used to demonstrate that the A(1) and A(O) domains are functionally coupled. This is the first description of an A(1) A(O) ATPase preparation in which the two domains (A(1) and A(O)) are fully conserved and functionally coupled.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | METHANOSARCINA-MAZEI GO1; CHEMIOSMOTIC ENERGY-CONVERSION; ARCHAEON HALOFERAX-VOLCANII; PLASMA-MEMBRANE ATPASE; ESCHERICHIA-COLI; METHANOGENIC ARCHAEA; SULFOLOBUS-ACIDOCALDARIUS; ACETOBACTERIUM-WOODII; ELECTRON-MICROSCOPY; F1F0-ATP SYNTHASE; A(1)A(0) ATPase; archaea; electron microscopy; functional coupling; purification |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 19 May 2021 06:32 |
| Last Modified: | 19 May 2021 06:32 |
| URI: | https://pred.uni-regensburg.de/id/eprint/38927 |
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