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Rapid affinity purification of retinal arrestin (48 kDa protein) via its light‐dependent binding to phosphorylated rhodopsin
Author(s) -
Wilden Ursula,
Wüst Eduard,
Weyand Ingo,
Kühn Hermann
Publication year - 1986
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(86)81507-1
Subject(s) - rhodopsin , arrestin , isoelectric focusing , retinal , affinity chromatography , chemistry , phosphorylation , phosphodiesterase , biochemistry , microbiology and biotechnology , chromatography , biology , receptor , enzyme , g protein coupled receptor
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.