Peng, Guohong and Fritzsch, Günter and Zickermann, Volker and Schaegger, Hermann and Mentele, Reinhardt and Lottspeich, Friedrich and Bostina, Mihnea and Radermacher, Michael and Huber, Robert and Stetter, Karl Otto and Michel, Hartmut (2003) Isolation, characterization and electron microscopic single particle analysis of the NADH : ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus. BIOCHEMISTRY, 42 (10). pp. 3032-3039. ISSN 0006-2960
Full text not available from this repository. (Request a copy)Abstract
The proton-translocating NADH:ubiquinone oxidoreductase (complex I) has been purified from Aquifex aeolicus, a hyperthermophilic eubacterium of known genome sequence. The purified detergent solubilized enzyme is highly active above 50 degreesC. The specific activity for electron transfer from NADH to decylubiquinone is 29 U/mg at 80 degreesC. The A. aeolicus complex I is completely sensitive to rotenone and 2-n-decyl-quinazoline-4-yl-amine. SDS polyacrylamide gel electrophoresis shows that it may contain up to 14 subunits. N-terminal amino acid sequencing of the bands indicates the presence of a stable subcomplex, which is composed of subunits E, F, and G. The isolated complex is highly stable and active in a temperature range from 50 to 90 degreesC, with a half-life of about 10 h at 80 degreesC. The activity shows a linear Arrhenius plot at 50-85 degreesC with an activation energy at 31.92 J/mol K. Single particle electron microscopy shows that the A. aeolicus complex I has the typical L-shape. However, visual inspection of averaged images reveals many more details in the external arm of the complex than has been observed for complex I from other sources. In addition, the angle (90degrees) between the cytoplasmic peripheral arm and the membrane intrinsic arm of the complex appears to be invariant.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ESCHERICHIA-COLI; MITOCHONDRIAL NADH; BACTERIAL; PROTEIN; PURIFICATION; TRANSLOCATION; DEHYDROGENASE; FLEXIBILITY; INHIBITORS; STABILITY; |
| 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: | 17 Aug 2021 13:58 |
| Last Modified: | 17 Aug 2021 13:58 |
| URI: | https://pred.uni-regensburg.de/id/eprint/39194 |
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