Voltage dependence of proton pumping by bacteriorhodopsin is regulated by the voltage-sensitive ratio of M1 to M2

Biophysical Journal
G NagelE Bamberg

Abstract

The voltage dependence of light-induced proton pumping was studied with bacteriorhodopsin (bR) from Halobacterium salinarum, expressed in the plasma membrane of oocytes from Xenopus laevis in the range -160 mV to +60 mV at different light intensities. Depending on the applied field, the quenching effect by blue light, which bypasses the normal photo and transport cycle, is drastically increased at inhibiting (negative) potentials, and is diminished at pump current increasing (positive) potentials. At any potential, two processes with different time constants for the M --> bR decay of approximately 5 ms (tau1) and approximately 20 ms (tau2) are obtained. At pump-inhibiting potentials, a third, long-lasting process with tau3 approximately 300 ms at neutral pH is observed. The fast processes (tau1, tau2) can be assigned to the decay of M2 in the normal pump cycle, i.e., to the reprotonation of the Schiff base via the cytoplasmic side, whereas tau3 is due to the decay of M1 without net pumping, i.e., the reprotonation of the Schiff base via the extracellular side. The results are supported by determination of photocurrents induced by bR on planar lipid films. The pH dependence of the slow decay of M1 is fully in agreement with the ...Continue Reading

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Citations

Oct 30, 2012·European Biophysics Journal : EBJ·Shizuma MiyazakiEiro Muneyuki
Sep 11, 2002·Journal of Molecular Biology·Thomas FriedrichErnst Bamberg
Jul 16, 2003·Journal of Molecular Biology·Silke Hippler-MreyenM Engelhard
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Nov 15, 2003·Proceedings of the National Academy of Sciences of the United States of America·Georg NagelErnst Bamberg
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