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J. Biochem, 1988, Vol. 103, No. 2 231-236
© 1988 Japanese Biochemical Society

research-article

DCCD-Sensitive Na+-Transport in the Membrane Vesicles of Halo bacterium halobium

Naoyuki Murakami and Tetsuya Konishi

Department of Radiochemistry-Biology, Niigata College of Pharmacy, Niigata Niigata 950-21

The effects of NN'-dicyclohexylcarbodiimide (DCCD) and various ionophores on light-induced 22Na+-transport were studied in right-side-out membrane vesicles, from Halobacterium halobium R1 M1. The light-induced Na+ efflux was inhibited at the same DCCD concentration (>40nmol/mg protein) as required for inhibition of the Na+-dependent membrane potential ({Delta}{psi}) formation. This supports our previous indication that the DCCD-sensitive, Na+-dependent transformation of pH-gradient ({Delta}pH) into {Delta}{Phi} is mediated by Na+/H+-antiporter (Murakami, N. and Konishi, T. (1985) J. Biochem. 98, 897–907). FCCP or a combination of valinomycin and triphenyltin (TPT) inhibits the light-induced Na+ efflux in accordance with the notion of protonmotive force ({Delta}µH)-driven antiporter. However, a marked lag in initiation of the Na+ efflux occurred in the presence of valinomycin, TPMP+, or a small amount of FCCP, suggesting that a gating step is involved in the Na+ efflux. On the other hand, the {Delta}pH-dissipating ionophore TPT did not cause the lag. A simultaneous determination of {Delta}{Phi}, {Delta}pH, and Na+ efflux rate at the initial stage of illumination revealed that the antiporter is gated by {Delta}{Phi} rather than {Delta}µH.


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