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Discovery of the light-driven sodium-motive pump Na + -rhodopsin (NaR) has initiated studies of the molecular mechanism of this novel membrane-linked energy transducer. In this paper, we investigated the photocycle of NaR from the marine flavobacterium  Dokdonia sp.  PRO95 and identified electrogenic and Na + -dependent steps of this cycle. We found that the NaR photocycle is composed of at least four steps:  NaR  519  +  hv  →  K  585  → ( L  450 ↔ M  495 ) →  O  585  →  NaR  519 . The third step is the only step that depends on the Na +  concentration inside right-side-out NaR-containing proteoliposomes, indicating that this step is coupled with Na +  binding to NaR. For steps 2, 3, and 4, the values of the rate constants are 4×10 4  s –1 , 4.7 × 10 3  M –1  s –1 , and 150 s –1 , respectively. These steps contributed 15, 15, and 70% of the total membrane electric potential (Δψ ~ 200 mV) generated by a single turnover of NaR incorporated into liposomes and attached to phospholipid-impregnated collodion film. On the basis of these observations, a mechanism of light-driven Na +  pumping by NaR is suggested.

Real-time kinetics of electrogenic Na+ transport by rhodopsin from the marine flavobacterium Dokdonia sp. PRO95