Flash-induced voltage changes in halorhodopsin from Natronobacterium pharaonis.

The flash-induced voltage response of halorhodopsin at high NaCl concentration comprises two main kinetic components. The first component with tau approximately 1 micros does not exceed 4% of the overall response amplitude and is probably associated with the formation of the L (hR520) intermediate. The second main component with tau approximately 1-2.5 ms which is independent of Cl- concentration can be ascribed to the transmembrane Cl- translocation during the L intermediate decay. The photoelectric response in the absence of Cl- has the opposite polarity and does not exceed 6% of the overall response amplitude at high NaCl concentration. A pH decrease results in substitution of the Cl(-)-dependent components by the photoresponse which is similar to that in the absence of Cl-. Thus, the difference between photoresponses of chloride-binding and chloride-free halorhodopsin forms resembles that of bacteriorhodopsin purple neutral and blue acid forms, respectively. The photovoltage data obtained can hardly be explained within the framework of the photocycle scheme suggested by Varo et al. [Biochemistry 34 (1995), 14490-14499]. We suppose that the O-type intermediate belongs to some form of halorhodopsin incapable of Cl- transport.