Phosphorylation of calmodulin by myosin light chain kinase is altered by exchange or duplication of EF-hand pairs.
We have previously demonstrated that under certain conditions, myosin light chain kinase can phosphorylate its activator, calmodulin. In this study we show that myosin light chain kinase from chicken gizzard can phosphorylate recombinant calmodulins in which the EF-hand pairs (Ca2+-binding domains) are duplicated or exchanged. Three mutants were used CaMNN (the amino-terminal EF-hand pair is duplicated), CaMCC (the carboxy-terminal EF-hand pair is duplicated) and CaMCN (the carboxy- and amino-terminal EF-hand pairs are switched). Myosin light chain kinase phosphorylated CaMNN and CaMCN to a greater extent than wild-type CaM but did not phosphorylate CaMCC. While CaMCC is a competitive inhibitor of myosin light chain kinase-catalyzed phosphorylation of myosin light chains, it did not prevent the phosphorylation of native calmodulin under the conditions employed in these studies. These data suggest that, although the carboxy- and amino-terminal EF-hand pairs are similar, their orientation can be distinguished by chicken gizzard myosin light chain kinase.
Davis
H W
HW
Department of Internal Medicine, University of Cincinnati Medical Center, Ohio 45267, USA.
D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002645:Chickens; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005895:Gizzard, Avian / Q000201:enzymology; D016296:Mutagenesis; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D013552:Swine
D002147:Calmodulin; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D009219:Myosin-Light-Chain Kinase; D002118:Calcium