Cooperativity and regulation of scallop myosin and myosin fragments.
Scallop heavy meromyosin (HMM) preparation obtained by a new improved method showed a Mg-ATPase activity that was activated 15-fold by calcium. The ATPase activity depended on ionic strength and reached maximum at 0.1 M without altering calcium sensitivity. The highly regulated HMM and myosin preparations showed cooperative properties not seen with unregulated subfragment 1 (S1). ATPase activity of myosin and HMM increased steeply with calcium concentration, yielding Hill coefficients about 3 and 4, respectively. Calcium binding by HMM and myosin became cooperative in the presence of ADP, AMP-PNP, or ADP.Vi yielding Hill coefficients of 1.8 and 2.8, respectively. Binding of calcium by HMM in the presence of ATP was also cooperative at physiological ionic strength, whereas at low ionic strength the data fit best to a simple binding curve. In contrast, calcium binding by unregulated S1 followed a normal binding curve and was not affected by the presence of nucleotide analogues. Calcium decreased the affinity of ADP and ADP-PNP to myosin and HMM, but had no effect on the nucleotide binding to S1. The results indicate that communication between the nucleotide and calcium binding sites requires the presence of two heads and exists only in the "off" state. We propose that in the presence of calcium, interaction between the two heads is disrupted and they act independently.
Kalabokis
V N
VN
Department of Biology, Brandeis University, Waltham, Massachusetts 02154-9110, USA.
Szent-Györgyi
A G
AG
D000244:Adenosine Diphosphate / Q000378:metabolism; D000266:Adenylyl Imidodiphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D008974:Mollusca; D015879:Myosin Subfragments / Q000378:metabolism*; D009218:Myosins / Q000378:metabolism*; D009994:Osmolar Concentration; D011485:Protein Binding
D015879:Myosin Subfragments; D000266:Adenylyl Imidodiphosphate; D000244:Adenosine Diphosphate; D009218:Myosins; D002118:Calcium