title: Two intermediates appear on the lumirhodopsin time scale after rhodopsin photoexcitation.
abstract: Absorbance difference spectra were recorded at 20 degrees C with a dense sequence of delay times from 1 to 128 micros after photolysis of lauryl maltoside suspensions of rhodopsin prepared from hypotonically washed bovine rod outer segments. Data were best fit by two-exponential components with a small, fast component (tau = 12 micros) occurring during the period that lumirhodopsin has been presumed to be stable. The shape of the spectral change corresponds to an approximately 2 nm red shift of the lumirhodopsin spectrum. Measurements with linearly polarized light verified that no absorbance changes associated with rotational diffusion were present in these preparations on this time scale, and experiments designed to enhance isorhodopsin production during photolysis showed no effect on the relative amplitude of the fast process. A similar process was previously observed in membrane suspensions of rhodopsin, but there the similarity of the change to rotational diffusion artifacts made conclusive identification of a second lumirhodopsin difficult. However, reexamination of polarized light measurements on rhodopsin in membrane supports the fact that the fast process seen here in detergent also takes place there. The new absorbance process occurs when time-resolved resonance Raman experiments have shown that the protonated Schiff base is moving from one hydrogen bond acceptor to another. The results are discussed in the context of possibly related processes on the same time scale that have been observed recently in artificial visual pigments with synthetic retinylidene chromophores and in a related rhodopsin mutant. The details of lumirhodopsin behavior are important because it is the last protonated Schiff base intermediate that occurs under physiological conditions.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Cruz 95064, USA.
authors.forename: Istvan
authors.initials: I
authors.lastname: Szundi
authors.forename: James W
authors.initials: JW
authors.lastname: Lewis
authors.forename: David S
authors.initials: DS
authors.lastname: Kliger
mesh_terms: D000818:Animals; D002417:Cattle; D007700:Kinetics; D008027:Light; D011487:Protein Conformation; D012243:Rhodopsin / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000378:metabolism* / Q000528:radiation effects; D012374:Rod Cell Outer Segment / Q000737:chemistry; D013053:Spectrophotometry
chemical_list: C018586:lumirhodopsin; D012243:Rhodopsin