Modulation of the phosphorylation and activity of calcium/calmodulin-dependent protein kinase II by zinc.
Calcium/calmodulin-dependent protein kinase II (CaMPK-II) is a key regulatory enzyme in living cells. Modulation of its activity, therefore, could have a major impact on many cellular processes. We found that Zn(2+) has multiple functional effects on CaMPK-II. Zn(2+) generated a Ca(2+)/CaM-independent activity that correlated with the autophosphorylation of Thr(286), inhibited Ca(2+)/CaM binding that correlated with the autophosphorylation of Thr(306), and inhibited CaMPK-II activity at high concentrations that correlated with the autophosphorylation of Ser(279). The relative level of autophosphorylation of these three sites was dependent on the concentration of zinc used. The autophosphorylation of at least these three sites, together with Zn(2+) binding, generated an increased mobility form of CaMPK-II on sodium dodecyl sulfate gels. Overall, autophosphorylation induced by Zn(2+) converts CaMPK-II into a different form than the binding of Ca(2+)/CaM. In certain nerve terminals, where Zn(2+) has been shown to play a neuromodulatory role and is present in high concentrations, Zn(2+) may turn CaMPK-II into a form that would be unable to respond to calcium signals.
Lengyel
I
I
Neuroscience Group, School of Biomedical Sciences, Faculty of Medicine and Health Sciences, University of Newcastle, Newcastle, New South Wales, Australia.
Fieuw-Makaroff
S
S
Hall
A L
AL
Sim
A T
AT
Rostas
J A
JA
Dunkley
P R
PR
D000818:Animals; D019921:Biotinylation; D019556:COS Cells; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism; D007700:Kinetics; D010446:Peptide Fragments / Q000737:chemistry; D010748:Phosphopeptides / Q000737:chemistry; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism; D016548:Prosencephalon / Q000201:enzymology; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D016704:Synapsins / Q000378:metabolism; D014162:Transfection; D019287:Zinc Sulfate / Q000494:pharmacology*
D002147:Calmodulin; D010446:Peptide Fragments; D010748:Phosphopeptides; D011994:Recombinant Proteins; D016704:Synapsins; D010768:Phosphoserine; D019287:Zinc Sulfate; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases