Photochemistry of the primary event in short-wavelength visual opsins at low temperature.
Two short-wavelength cone opsins, frog (Xenopus laevis) violet and mouse UV, were expressed in mammalian COS1 cells, purified in delipidated form, and studied using cryogenic UV-vis spectrophotometry. At room temperature, the X. laevis violet opsin has an absorption maximum at 426 nm when generated with 11-cis-retinal and an absorption maximum of 415 nm when generated with 9-cis-retinal. The frog short-wavelength opsin has two different batho intermediates, one stable at 30 K (lambda(max) approximately 446 nm) and the other at 70 K (lambda(max) approximately 475 nm). Chloride ions do not affect the absorption maximum of the violet opsin. At room temperature, mouse UV opsin has an absorption maximum of 357 nm, while at 70 K, the pigment exhibits a bathochromic shift to 403 nm with distinct vibronic structure and a strong secondary vibronic band at 380 nm. We have observed linear relationships when analyzing the energy difference between the initial and bathochromic intermediates and the normalized difference spectra of the batho-shifted intermediates of rod and cone opsins. We conclude that the binding sites of these pigments change from red to green to violet via systematic shifts in the position of the primary counterion relative to the protonated Schiff base. The mouse UV cone opsin does not fit this trend, and we conclude that wavelength selection in this pigment must operate via a different molecular mechanism. We discuss the possibility that the mouse UV chromophore is initially unprotonated.
Vought
B W
BW
Department of Chemistry, Syracuse University, New York 13244-4100, USA.
Dukkipatti
A
A
Max
M
M
Knox
B E
BE
Birge
R R
RR
D000042:Absorption; D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002417:Cattle; D002712:Chlorides / Q000737:chemistry; D003080:Cold Temperature; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008969:Molecular Sequence Data; D010777:Photochemistry; D011489:Protein Denaturation; D017949:Retinal Cone Photoreceptor Cells / Q000737:chemistry; D012172:Retinaldehyde / Q000737:chemistry; D012243:Rhodopsin / Q000737:chemistry; D017299:Rod Opsins / Q000737:chemistry* / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D013464:Sulfuric Acids; D014466:Ultraviolet Rays; D014982:Xenopus laevis
D002712:Chlorides; D017299:Rod Opsins; D013464:Sulfuric Acids; C095853:short-wavelength opsin; D012243:Rhodopsin; C033158:sulfuric acid; D012172:Retinaldehyde