title: Regeneration of bovine and octopus opsins in situ with natural and artificial retinals.
abstract: We consider the problem of color regulation in visual pigments for both bovine rhodopsin (lambda max = 500 nm) and octopus rhodopsin (lambda max = 475 nm). Both pigments have 11-cis-retinal (lambda max = 379 nm, in ethanol) as their chromophore. These rhodopsins were bleached in their native membranes, and the opsins were regenerated with natural and artificial chromophores. Both bovine and octopus opsins were regenerated with the 9-cis- and 11-cis-retinal isomers, but the octopus opsin was additionally regenerated with the 13-cis and all-trans isomers. Titration of the octopus opsin with 11-cis-retinal gave an extinction coefficient for octopus rhodopsin of 27,000 +/- 3000 M-1 cm-1 at 475 nm. The absorption maxima of bovine artificial pigments formed by regenerating opsin with the 11-cis dihydro series of chromophores support a color regulation model for bovine rhodopsin in which the chromophore-binding site of the protein has two negative charges: one directly hydrogen bonded to the Schiff base nitrogen and another near carbon-13. Formation of octopus artificial pigments with both all-trans and 11-cis dihydro chromophores leads to a similar model for octopus rhodopsin and metarhodopsin: there are two negative charges in the chromophore-binding site, one directly hydrogen bonded to the Schiff base nitrogen and a second near carbon-13. The interaction of this second charge with the chromophore in octopus rhodopsin is weaker than in bovine, while in metarhodopsin it is as strong as in bovine.
authors.affiliation: Department of Physiology and Biophysics, University of Illinois, Urbana 61801.
authors.forename: Y
authors.initials: Y
authors.lastname: Koutalos
authors.forename: T G
authors.initials: TG
authors.lastname: Ebrey
authors.forename: M
authors.initials: M
authors.lastname: Tsuda
authors.forename: K
authors.initials: K
authors.lastname: Odashima
authors.forename: T
authors.initials: T
authors.lastname: Lien
authors.forename: M H
authors.initials: MH
authors.lastname: Park
authors.forename: N
authors.initials: N
authors.lastname: Shimizu
authors.forename: F
authors.initials: F
authors.lastname: Derguini
authors.forename: K
authors.initials: K
authors.lastname: Nakanishi
authors.forename: H R
authors.initials: HR
authors.lastname: Gilson
mesh_terms: D000818:Animals; D002417:Cattle; D005136:Eye Proteins / Q000378:metabolism*; D007700:Kinetics; D049831:Octopodiformes; D012168:Retinal Pigments / Q000378:metabolism*; D012172:Retinaldehyde / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000378:metabolism*; D012176:Retinoids / Q000378:metabolism*; D012243:Rhodopsin / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism; D017299:Rod Opsins; D013053:Spectrophotometry / Q000379:methods; D013329:Structure-Activity Relationship
chemical_list: D005136:Eye Proteins; D012168:Retinal Pigments; D012176:Retinoids; D017299:Rod Opsins; D012243:Rhodopsin; D012172:Retinaldehyde