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Spectral properties of the rhod opsin system of the crayfish astacus leptodactylus


, : Spectral properties of the rhod opsin system of the crayfish astacus leptodactylus. Photochemistry & Photobiology 39(3): 379-390

The absorption spectra of the membrane-bound and of the digitonin-solubilized visual pigment of crayfish A. leptodactylus were investigated by conventional spectrophotometry. A method was developed to isolate purified rhabdoms almost entirely free from screening pigments from a single retina. The quantity of isolated and purified rhabdoms from a single retina was sufficient to measure the absorption spectra of the visual pigment. The absorption spectra of the chromoprotein system (R and M) show that the membrane-bound and the digitonin-solubilized visual pigment isomers are stable at 0.degree. C and pH 7.0. Rhodopsin and metarhodopsin are photoreversible under these conditions without any light-induced denaturation. The difference spectra for the chromoprotein isomers and those of different photostationary states yield maximal values for .DELTA.E at 570 and 485 nm. At neutral pH, 0.degree. C, .lambda.max of rhodopsin is 530 nm. Irradiation with light of .lambda. = 630 to 640 nm isomerizes rhodopsin nearly quantitatively to metarhodopsin with .lambda.max, of 500 nm. The molar extinction coefficient of metarhodopsin is greater than that of rhodopsin by a factor of .apprx. 1.41, each measured at its respective .lambda.max. Metarhodopsin can be isomerized to rhodopsin by irradiating at .lambda. < 630 nm. As the absorption spectra of the 2 chromoprotein isomers overlap, only part of the metarhodopsin can be reversed to rhodopsin. The maximal photoreversion can be achieved by irradiating at 460 nm. The stability of the digitonin-solubilized chromoprotein is remarkably dependent on temperature. Warming the digitonin extract of rhabdoms from 0.degree.-20.degree. or 30.degree. C caused a shift of the rhodopsin spectrum to shorter wavelengths (.lambda.max = 485 nm) accompanied by a decrease of E.lambda.maX by .apprx. 30%.

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