Squid visual arrestin: cDNA cloning and calcium-dependent phosphorylation by rhodopsin kinase (SQRK).
Arrestin binding to rhodopsin is one of the major mechanisms of termination of photoresponses in both vertebrates and invertebrates. Here we report the cDNA cloning and characterization of a 48-kDa visual arrestin from squid (Loligo pealei). The cDNA encoded a protein that had 56-64% amino acid sequence similarity to reported arrestin sequences. This protein does not encode any distinct modular domains but contains five fingerprint regions that have been identified within arrestins. Antibodies raised to the recombinant arrestin protein detected arrestin expression only in the eye and recognized a doublet in photoreceptor membranes, representing unphosphorylated and phosphorylated arrestin. In squid eye membranes, arrestin was phosphorylated in a Ca2+-dependent manner and this phosphorylation was inhibited by antibodies raised against squid rhodopsin kinase, but not by inhibitors of protein kinase C or calmodulin kinase. Addition of purified squid rhodopsin kinase to washed rhabdomeric membranes resulted in phosphorylation of rhodopsin, and arrestin was also phosphorylated when calcium was present. This is the first report of a rhodopsin kinase phosphorylating an arrestin substrate, and suggests a dual role for this kinase in the inactivation of the squid visual system.
Mayeenuddin
Linnia H
LH
Department of Pharmacology, University of Toronto, Toronto, Canada.
Mitchell
Jane
J
D000595:Amino Acid Sequence; D000818:Animals; D019393:Arrestin / Q000235:genetics* / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D003001:Cloning, Molecular; D049832:Decapodiformes; D005136:Eye Proteins; D051606:G-Protein-Coupled Receptor Kinase 1; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D019393:Arrestin; D005136:Eye Proteins; C474322:visual arrestin protein, Loligo pealei; D011494:Protein Kinases; D051606:G-Protein-Coupled Receptor Kinase 1; D002118:Calcium