Abstract
The rotenone sensitive NADH: menaquinone oxidoreductase (NDH-I or complex I) from the thermohalophilic bacterium Rhodothermus marinus has been purified and characterized. Three of its subunits react with antibodies against 78, 51, and 21.3c kDa subunits of Neurospora crassa complex I. The optimum conditions for NADH dehydrogenase activity are 50°C and pH 8.1, and the enzyme presents a K M of 9 μM for NADH. The enzyme also displays NADH:quinone oxidoreductase activity with two menaquinone analogs, 1,4-naphtoquinone (NQ) and 2,3-dimethyl-1,4-naphtoquinone (DMN), being the last one rotenone sensitive, indicating the complex integrity as purified. When incorporated in liposomes, a stimulation of the NADH:DMN oxidoreductase activity is observed by dissipation of the membrane potential, upon addition of CCCP. The purified enzyme contains 13.5 ± 3.5 iron atoms and ∼3.7 menaquinone per FMN. At least five iron—sulfur centers are observed by EPR spectroscopy: two [2Fe–2S]2+/1+ and three [4Fe–4S]2+/1+ centers. By fluorescence spectroscopy a still unidentified chromophore was detected in R. marinus complex I.
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Fernandes, A.S., Pereira, M.M. & Teixeira, M. Purification and Characterization of the Complex I from the Respiratory Chain of Rhodothermus marinus . J Bioenerg Biomembr 34, 413–421 (2002). https://doi.org/10.1023/A:1022509907553
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DOI: https://doi.org/10.1023/A:1022509907553