Obtaining and characterization of EF-hand mutants of recoverin.
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.
Alekseev
A M
AM
Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Puschino, Moscow Region, Russia.
Shulga-Morskoy
S V
SV
Zinchenko
D V
DV
Shulga-Morskaya
S A
SA
Suchkov
D V
DV
Vaganova
S A
SA
Senin
I I
II
Zargarov
A A
AA
Lipkin
V M
VM
Akhtar
M
M
Philippov
P P
PP
D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D002417:Cattle; D005136:Eye Proteins; D051606:G-Protein-Coupled Receptor Kinase 1; D051597:Hippocalcin; D008074:Lipoproteins; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009419:Nerve Tissue Proteins; D010641:Phenotype; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D017433:Protein Structure, Secondary; D051603:Recoverin; D012160:Retina; D012374:Rod Cell Outer Segment / Q000378:metabolism*
D002135:Calcium-Binding Proteins; D005136:Eye Proteins; D008074:Lipoproteins; D009419:Nerve Tissue Proteins; D047428:Protein Kinase Inhibitors; D051603:Recoverin; D051597:Hippocalcin; D011494:Protein Kinases; D051606:G-Protein-Coupled Receptor Kinase 1; D002118:Calcium