Bacteriorhodopsins containing modified chromophores: A study on the wild type and D96N mutant of Halobacterium salinarum |
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Authors: | L. V. Khitrina E. V. Mironova A. A. Khodonov |
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Affiliation: | (1) Belozersky Institute of Physico-Chemical Biology, Moscow Lomonosov State University, Moscow, 119991, Russia;(2) Lomonosov State Academy of Fine Chemical Technology, prosp. Vernadskogo, 86, Moscow, 117571, Russia |
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Abstract: | Modification of the chromophore in bacteriorhodopsin (BR) from ET1001 and D96N strains of Halobacterium salinarum (halobium) was carried out. Purple membranes were decolored by means of light-dependent hydroxylaminolysis. The all-trans -isomers of retinal and its 3,4-didehydro-, 4-keto-, and phenyl analogs were reconstituted into apomembranes. Absorption maxima of the homonymic pigments in both strains were similar. The kinetics of the M-intermediates in the mode of a single turnover of the photocycle induced by a short light flash (532 nm, 15 ns) was compared. For the investigated bacteriorhodopsin analogs the efficiency of the M-intermediate formation did not exhibit any reliable dependence on the point mutation. Both for ET1001 and for D96N strains the M-relaxation of the 4-ketoBR was distinctly biphasic, with the slow phase comprising about 10–15% of the signal amplitude. Replacement of the ionone ring by phenyl caused a weak deceleration of the M relaxation (~1.5-fold decrease in t 1/2). Independence of the photocycle deceleration of the point mutation and chromophore modification was shown for all BR analogs studied. |
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Keywords: | bacteriorhodopsin M-intermediate D96N apomembranes 4-ketobacteriorhodopsin 3,4-didehydrobacteriorhodopsin phenylbacteriorhodopsin retinal analogs |
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