Binding property of avian skeletal muscle ryanodine receptor isoforms with dihydropyridine receptor and calmodulin.
Ca(2+) release during excitation-contraction coupling in avian skeletal muscle is controlled by two ryanodine receptor isoforms, alphaRYR and betaRYR. Two other proteins, dihydropyridine receptor (DHPR) and calmodulin (CaM), have been shown to play important roles in regulating the RYR channel activity. In the current study, we measured the protein contents of DHPR and RYR in turkey skeletal muscle and obtained a ratio of 1:1 between DHPR and alphaRYR which suggests that only a subpopulation of alphaRYR is interacting with DHPR. Two CaM derivatives, the photoactivable crosslinking probe [(125)I]-Bz-CaM and metabolically labeled probe [(35)S]CaM, were used to study the interaction between CaM and RYR isoforms in turkey skeletal muscle. The alphaRYR and betaRYR displayed a marked difference in their CaM binding behavior. At a Ca(2+) concentration of 200 microM, CaM bound to both isoforms at a ratio of one CaM molecule per one RYR subunit. At a Ca(2+) concentration of <10 nM, CaM bound primarily to alphaRYR and the binding affinity was significantly lower than that at micromolar level of Ca(2+) concentration. Cloning and sequencing of putative CaM binding sites in alphaRYR and betaRYR suggests that differences in primary structures of the CaM binding sites of each RYR isoform may contribute to the differential CaM binding behavior of alphaRYR and betaRYR.
Chiang
Wen
W
Department of Food Science and Human Nutrition, Michigan State University, R3365 Anthony Hall, East Lansing, MI 48824, USA. chiangw1@msu.edu
Byrem
Todd
T
Zhang
Haiyan
H
Strasburg
Gale
G
D000818:Animals; D002118:Calcium / Q000378:metabolism; D020746:Calcium Channels, L-Type / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D019837:Ryanodine Receptor Calcium Release Channel / Q000737:chemistry / Q000378:metabolism*; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D016415:Sequence Alignment; D014421:Turkey
D020746:Calcium Channels, L-Type; D002147:Calmodulin; D020033:Protein Isoforms; D019837:Ryanodine Receptor Calcium Release Channel; D002118:Calcium