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Obtaining and characterization of EF-hand mutants of recoverin
A M Alekseev1, S V Shulga-Morskoy, D V Zinchenko
1Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Puschino, Moscow Region, Russia.
FEBS Letters
|December 23, 1998
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.