Regulation of phototransduction in short-wavelength cone visual pigments via the retinylidene Schiff base counterion.
Short-wavelength visual pigments (SWS1) have lambda(max) values that range from the ultraviolet to the blue. Like all visual pigments, this class has an 11-cis-retinal chromophore attached through a Schiff base linkage to a lysine residue of opsin apoprotein. We have characterized a series of site-specific mutants at a conserved acidic residue in transmembrane helix 3 in the Xenopus short-wavelength sensitive cone opsin (VCOP, lambda(max) approximately 427 nm). We report the identification of D108 as the counterion to the protonated retinylidene Schiff base. This residue regulates the pK(a) of the Schiff base and, neutralizing this charge, converts the violet sensitive pigment into one that absorbs maximally in the ultraviolet region. Changes to this position cause the pigment to exhibit two chromophore absorbance bands, a major band with a lambda(max) of approximately 352-372 nm and a minor, broad shoulder centered around 480 nm. The behavior of these two absorbance bands suggests that these represent unprotonated and protonated Schiff base forms of the pigment. The D108A mutant does not activate bovine rod transducin in the dark but has a significantly prolonged lifetime of the active MetaII state. The data suggest that in short-wavelength sensitive cone visual pigments, the counterion is necessary for the characteristic rapid production and decay of the active MetaII state.
Babu
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Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University, 766 Irving Avenue, Syracuse, New York 13210, USA.
Dukkipati
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Birge
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Knox
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D000818:Animals; D001224:Aspartic Acid / Q000235:genetics; D019556:COS Cells; D002417:Cattle; D018698:Glutamic Acid / Q000235:genetics; D005973:Glutamine / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D011522:Protons; D017949:Retinal Cone Photoreceptor Cells / Q000737:chemistry* / Q000378:metabolism; D012172:Retinaldehyde / Q000737:chemistry / Q000378:metabolism; D012176:Retinoids / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017299:Rod Opsins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012545:Schiff Bases / Q000737:chemistry / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D055672:Static Electricity; D014785:Vision, Ocular / Q000235:genetics; D014981:Xenopus
D011522:Protons; D012176:Retinoids; D017299:Rod Opsins; D012545:Schiff Bases; C121835:retinylidene chromophore; C095853:short-wavelength opsin; D005973:Glutamine; D001224:Aspartic Acid; D018698:Glutamic Acid; D012172:Retinaldehyde