1. Obtaining and characterization of EF-hand mutants of recoverin.
- Author
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Alekseev AM, Shulga-Morskoy SV, Zinchenko DV, Shulga-Morskaya SA, Suchkov DV, Vaganova SA, Senin II, Zargarov AA, Lipkin VM, Akhtar M, and Philippov PP
- Subjects
- Animals, Binding Sites, Calcium metabolism, Calcium-Binding Proteins chemistry, Cattle, G-Protein-Coupled Receptor Kinase 1, Hippocalcin, Mutagenesis, Site-Directed, Phenotype, Phosphorylation, Protein Structure, Secondary, Recoverin, Retina, Calcium-Binding Proteins genetics, Calcium-Binding Proteins metabolism, Eye Proteins, Lipoproteins, Mutation, Nerve Tissue Proteins, Protein Kinase Inhibitors, Protein Kinases, Rod Cell Outer Segment metabolism
- Abstract
Several EF-hand recoverin mutants were obtained and their abilities to bind to photoreceptor membranes and to inhibit rhodopsin kinase were determined. The mutants with the 'spoiled' 2nd, 3rd or (2nd+3rd) EF-hand structures did not act upon the kinase activity in the microM range of Ca2+ concentrations. Mutations of the 4th EF hand, which 'repaired' its Ca2+-binding activity, resulted in recoverin with three 'working' Ca2+-binding sites. The latter mutant inhibited rhodopsin kinase even more effectively than the wild-type recoverin, containing two working Ca2+-binding structures.
- Published
- 1998
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