| Abstract: | Quantitative action spectroscopy was performed in Halobacterium halobium. using four suited pigment mutants, namely the bacteriorhodopsin and halorhodopsin positive mutant strain M-l (BR+, HR+), the bacteriorhodopsin positive but halorhodopsin negative strain M-18 (BR+, HR-), the bacteriorhodopsin negative but halorhodopsin positive strain L-33 (BR-, HR+), and the bacteriorhodopsin and halorhodopsin negative strain L-07 (BR-, HR+). The approached questions were: First, photoenergetic synergism of halorhodopsin and bacteriorhodopsin in intact cells; second, photochromism and cellular function of the blue light-absorbing intermediates, i.e. M-412 and HR-410 in bacteriorhodopsin and in halorhodopsin, respectively. Dark-adapted cells of mutant strain M-l show wavelength-dependency of quantum yield of photo-phosphorylation, φATP. An 1.4-fold enhancement was found at 575 nm wavelength where the long wavelength absorbance bands of bacteriorhodopsin and halorhodopsin intersect. The enhancement vanished after a 30 min pulse of orange light (600 Wm-2 bandpass from 495 to 750 nm), but was restored after a 30 min pulse of blue light (100 Wm-2 bandpass from 325 to 480 nm). Photoreversibility of this enhancement probably reflects phototransformation of halorhodopsin from its ground state into its inactive intermediate, HR-410, and vice versa. The halorhodopsin-mediated enhancement with maximum quantum yield of photophosphorylation, φATP= 0.06, i.e. a quantum requirement of = 17 photons/ATP, is partly substituted by a rise in phosphate potential and explained in terms of a voltage-regulated gating effect on the H+-driven ATP-synthase, superimposed on the chemiosmotic mechanism of energy coupling. The blue-absorbing photochromic intermediate, M-412 of bacteriorhodopsin, dissipates light energy upon photoexcitation that is reflected by a spectral decline in quantum yield of photophosphorylation to a minimum value of = 0.01 at 415 nm, i.e. a quantum requirement of = 100 photons/ATP. |
