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The requirement for Na+, however, is greatly reduced in the presence of 400 m M K + (curve 1 in Fig. 8 mM. The cooperative effect of K+ or Mg 2 + with Na + is important in considering the respiratory activity of intact cells. Since V. alginolyticus normally contains 400 mM K+ and less than 100 mM Na+, cellular K + together with Mg24* plays a critical role in the maintenance of respiratory activity when the cellular Na + decreases to a low level. Rb+ and C s + behave in a similar way to K + , but Li + rather inhibits the effect of Na + .

C. DISCOVERY OF RESPIRATION-DRIVEN SODIUM PUMP In 1981, Tokuda and Unemoto found that V. alginolyticus is able to generate membrane potential (Δψ) at alkaline pH even in the presence of a proton conductor, carbonyl cyanide m-chlorophenylhydrazone (CCCP). Although the Δψ gen- 2 . 5 was unaffected by up to 50 μΜ CCCP (Tokuda and Unemoto, 1982). Importantly, the generation of CCCP-resistant Δψ was dependent on respiration and was always accompanied by the accumulation of H+ inside the cells, leading to the generation of ΔρΗ (acidic inside) of similar magnitude to that of Δψ.

And Schatz, G. (1980). A two-subunit cytochrome c oxidase (cytochrome aa3) from Paracoccus denitrificans. Proc. Natl. Acad. Sei. A. 77, 196-200. , Power, D. , and Thomas, E. W. (1980). Purification of an NADH: Dichlorophenolindophenol oxidoreductase from Bacillus stearothermophilus. Biochem. J. 191, 457-465. Margulis, L. (1970). " Yale University Press, New Haven, Connecticut. , and Ameyama, M. (1982). o-type cytochrome oxidase in the membrane of aerobically grown Pseudomonas aeruginosa. FEBS Lett.

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