Rhodopsin kinase: studies on the sequence of and the recognition motif for multiphosphorylations.
Peptides of 10-12 amino acids in length, which overlapped with the sequence of the last 20 amino acids in the C-terminal tail of rhodopsin, were synthesized and used as substrates for rhodopsin kinase. In all cases the phosphorylation of the peptides was found to be greatly stimulated (> 20-fold) by the presence of light-activated rhodopsin (Rho*). The incorporation of 32P at seven Ser/Thr residues that are the potential sites of phosphorylation was quantified, and the results were analyzed in terms of two parameters. First, a global comparison of phosphorylation at each site was made when the propensity for the modification was found to be in the order: Ser 343 > Ser 338 > Thr 336 > Ser 334, Thr 342 > Thr 335, Thr 340. Second, the peptides were aligned on a hypothetical template with the residue to be phosphorylated occupying the P-position, and the manner in which the nature of the surrounding residues effected the phosphorylation was assessed. It was found that the optimal phosphorylation of the P-site Ser/Thr occurs if it has at least one residue on the amino side and five on the acyl side and also contains a neutral residue, preferably small (A, P, S, T) at the P+4 position.(ABSTRACT TRUNCATED AT 250 WORDS)
Pullen
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N
Department of Biochemistry, University of Southampton, United Kingdom.
Akhtar
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D005136:Eye Proteins; D051606:G-Protein-Coupled Receptor Kinase 1; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010748:Phosphopeptides / Q000737:chemistry; D010768:Phosphoserine / Q000378:metabolism; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D012243:Rhodopsin / Q000737:chemistry* / Q000378:metabolism; D012374:Rod Cell Outer Segment / Q000737:chemistry; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D005136:Eye Proteins; D010748:Phosphopeptides; D010768:Phosphoserine; D012243:Rhodopsin; D011494:Protein Kinases; D051606:G-Protein-Coupled Receptor Kinase 1