The presence of H+ and Na+-translocating ATPases in Methanobacterium thermoautotrophicum and their possible function under alkaline conditions

AbstractTwo ATPases with different apparent molecular masses of approx. 500 kDa and 400 kDa were identified in the EDTA extract of the cell membranes of Methanobacterium thermoautotrophicum. Western blotting with polyclonal antiserum reactive with β-subunit of mitochondrial ATPase from rat liver and yeast was used for further analysis of these ATPases. A strong crossreactivity with a single protein band with an apparent molecular weight of about 53 kDa (similar to β-subunit of F-type ATPase from other sources) was found in protein extracts of whole cells of Methanobacterium thermoautotrophicum strains ΔH and Marburg, as well as of Methanospirillum hungatei. This indicates the presence of F-type ATPase in methanogens. ATP synthesis driven by membrane potential which was generated by artificially-imposed ΔpH in the presence of protonophorous uncoupler and sodium ions was stimulated by bafilomycin A1, an inhibitor of V- and A-type ATPases, as well as by harmaline, an inhibitor of Na+/H+ antiporter. These results indicate that cells of Methanobacterium thermoautotrophicum strain ΔH contain the F-type ATP synthase which is Na+-translocating in addition to V- or A-type ATP synthase which is H+-translocating