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.
Department of Molecular and Cellular Pharmacology, University of Miami School of Medicine, Florida 33136, USA.
D K
DK
Satpaev
C K
CK
Chen
A
A
Scotti
M I
MI
Simon
J B
JB
Hurley
V Z
VZ
Slepak
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