Premium
TIME‐RESOLVED FLUOROMETRY OF BACTERIORHODOPSIN
Author(s) -
Sherman Warren V.
Publication year - 1982
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1982.tb04403.x
Subject(s) - bacteriorhodopsin , chromophore , rhodopsin , chemistry , fluorescence , retinal , photoprotein , tryptophan , kinetics , retinaldehyde , photochemistry , fluorescence spectroscopy , biophysics , crystallography , membrane , biochemistry , amino acid , optics , biology , bioluminescence , physics , quantum mechanics
A mode‐locked Nd:YAG laser was used to excite the aromatic amino acid residues of bacteriorhodopsin in the purple membrane and the tryptophan (Trp) fluorescence decay analyzed with a streak camera (λ> 380 nm). The decay kinetics are resolvable into two first‐order half‐times (1.5 and 0.17 ns, respectively), while for retinylidene‐free bacterioopsin, only the longer‐lived Trp emission was observed. The shorter‐lived species reappeared upon regeneration of bacteriorhodopsin by addition of retinal to bacterioopsin but not on treatment of the latter with an equivalent of retinol. It is proposed that these results are consistent with a structural model in which the 7‐8 Trp's distributed among sections A, C, E and F of the seven helical segments A‐G of native bacteriorhodopsin are distinguishable by their distances from the chromophore. Assuming a Förster mechanism for energy transfer with R o = 25 and 32 Å, respectively, for retinylidene chromophore and retinol the Trp's may be divided into two groups: (i) those completely quenched by retinol and partly quenched by retinal (τ= 0.17) with R ≃ 18 Å and (ii) those (τ= 1.5 ns) which are quenched neither by chromophore nor retinol with R > ca. 30 Å. These results are consistent with and support some of the best models of Engelman et al. (1980) for the protein conformation in the purple membrane.