Rapid affinity purification of retinal arrestin (48 kDa protein) via its light-dependent binding to phosphorylated rhodopsin.
Arrestin (also named '48 kDa protein' or 'S-antigen') is a soluble protein involved in controlling light-dependent cGMP phosphodiesterase activity in retinal rods, and is also known for its ability to induce autoimmune uveitis of the eye. We report a rapid and simple purification method based on the property of arrestin to bind specifically and reversibly to illuminated and phosphorylated rhodopsin [(1984) FEBS Lett. 176, 473-478]. This method does not require column chromatography and yields about 2-4 mg purified arrestin from 15 bovine retinas. Pure arrestin can be resolved by isoelectric focusing into at least 10 distinct bands, all of which stain with a monoclonal antibody specific for S-antigen.
D000818:Animals; D000941:Antigens / Q000302:isolation & purification* / Q000378:metabolism; D019393:Arrestin; D002417:Cattle; D002498:Centrifugation; D002845:Chromatography; D002851:Chromatography, High Pressure Liquid; D005136:Eye Proteins / Q000302:isolation & purification* / Q000378:metabolism; D007525:Isoelectric Focusing; D008027:Light; D010766:Phosphorylation; D012160:Retina / Q000032:analysis*; D012168:Retinal Pigments / Q000378:metabolism*; D012243:Rhodopsin / Q000378:metabolism* / Q000528:radiation effects
D000941:Antigens; D019393:Arrestin; D005136:Eye Proteins; D012168:Retinal Pigments; D012243:Rhodopsin