title: Purification and characterization of bovine cone arrestin (cArr).
abstract: To elucidate the quenching mechanism of phototransduction in vertebrate cone photoreceptors, a cDNA clone encoding cone specific arrestin (cArr) was isolated from a bovine retinal cDNA library using a human cArr cDNA probe. Affinity-purified anti-peptide antibody specific to cArr was prepared. Immunohistochemical staining displayed specific labeling of cArr in cone photoreceptors and immunoblotting identified a 46 kDa protein band. We purified cArr from bovine retinas by sequential column chromatography using DEAE-cellulose, gel filtration and mono Q columns. Binding studies revealed no binding of cArr to rhodopsin regardless of whether it was bleached and/or phosphorylated. cArr also failed to bind to heparin-Sepharose under conditions which rod arrestin (rArr) bound to the column. The present data suggest that cArr may play a role in the quenching of phototransduction in cone photoreceptors and that its activity therein is different to that of rArr.
authors.affiliation: Department of Ophthalmology, Sapporo Medical University School of Medicine, S-1 W-16, Chuo-ku, Sapporo, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Maeda
authors.forename: H
authors.initials: H
authors.lastname: Ohguro
authors.forename: H
authors.initials: H
authors.lastname: Sohma
authors.forename: Y
authors.initials: Y
authors.lastname: Kuroki
authors.forename: H
authors.initials: H
authors.lastname: Wada
authors.forename: S
authors.initials: S
authors.lastname: Okisaka
authors.forename: A
authors.initials: A
authors.lastname: Murakami
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019393:Arrestin / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D002417:Cattle; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D006493:Heparin / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010766:Phosphorylation; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017949:Retinal Cone Photoreceptor Cells / Q000737:chemistry* / Q000166:cytology; D012243:Rhodopsin / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D019393:Arrestin; D011994:Recombinant Proteins; D006493:Heparin; D012243:Rhodopsin