All- trans retinal constitutes the functional chromophore in Chlamydomonas rhodopsin

• Published in: Biophysical journal Vol. 60; no. 6; pp. 1477 - 1489
• Main Authors: Hegemann, P., Gärtner, W., Uhl, R.
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Elsevier Inc 01.12.1991; Biophysical Society
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Non metallic chromoproteins, photoproteins; Proteins; Rhodopsin; Algae; Chlorophycophyta; Photoprotein; Chlamydomonas; Isomer; In vivo; Phototaxis; Analog; Reconstituted system; Retinal; Thallophyta; Chromophore
• ISSN: 0006-3495; 1542-0086
• DOI: 10.1016/S0006-3495(91)82183-X

Orientation of the green alga Chlamydomonas in light (phototaxis and stop responses) is controlled by a visual system with a rhodopsin as the functional photoreceptor. Here, we present evidence that in Chlamydomonas wild-type cells all- trans retinal is the predominant isomer and that it is present in amounts similar to that of the rhodopsin itself. The ability of different retinal isomers and analog compounds to restore photosensitivity in blind Chlamydomonas cells (strain CC2359) was tested by means of flash-induced light scattering transients or by measuring phototaxis in a taxigraph. All- trans retinal reconstitutes behavioral light responses within one minute, whereas cis-isomers require at least 50 × longer incubation times, suggesting that the retinal binding site is specific for all- trans retinal. Experiments with 13-demethyl(dm)-retinal and short-chained analogs reveal that only chromophores with a β-methyl group and at least three double bonds in conjugation with the aldehyde mediate function. Because neither 13-dm-retinal, nor 9,12-phenylretinal restores a functional rhodopsin, a trans/13- cis isomerisation seems to take place in the course of the activation mechanism. We conclude that with respect to its chromophore, Chlamydomonas rhodopsin bears a closer resemblence to bacterial rhodopsins than to visual rhodopsins of higher animals.