Autophosphorylation and ADP regulate the Ca2+-dependent interaction of recoverin with rhodopsin kinase.
Recoverin is a 23 kDa myristoylated Ca2+-binding protein that inhibits rhodopsin kinase. We have used surface plasmon resonance to investigate the influences of Ca2+, myristoylation, and adenine nucleotides on the recoverin-rhodopsin kinase interaction. Our analyses confirmed that Ca2+ is required for recoverin to bind RK. Myristoylation had little effect on the affinity of recoverin for the kinase, but it raised the K0.5 for Ca2+ from 150 nM for nonacylated recoverin to 400 nM for myristoylated recoverin. Finally, our studies also revealed two separate and previously unreported effects of adenine nucleotides on the recoverin-rhodopsin kinase binding. The interaction is weakened by autophosphorylation of the kinase, and it is strengthened by the presence of ADP.
Satpaev
D K
DK
Department of Molecular and Cellular Pharmacology, University of Miami School of Medicine, Florida 33136, USA.
Chen
C K
CK
Scotti
A
A
Simon
M I
MI
Hurley
J B
JB
Slepak
V Z
VZ
D000244:Adenosine Diphosphate / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors / Q000378:metabolism; D000495:Allosteric Site; D015374:Biosensing Techniques; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D005136:Eye Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D051606:G-Protein-Coupled Receptor Kinase 1; D051597:Hippocalcin; D007700:Kinetics; D008074:Lipoproteins; D019814:Myristic Acid / Q000378:metabolism; D009419:Nerve Tissue Proteins; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D051603:Recoverin; D013379:Substrate Specificity
D002135:Calcium-Binding Proteins; D005136:Eye Proteins; D008074:Lipoproteins; D009419:Nerve Tissue Proteins; D047428:Protein Kinase Inhibitors; D019814:Myristic Acid; D051603:Recoverin; D051597:Hippocalcin; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D051606:G-Protein-Coupled Receptor Kinase 1; D010749:Phosphoprotein Phosphatases; D002118:Calcium