Actin-induced local conformational change in the myosin molecule. I. Effect of metal ions and nucleotides on the conformational change around a specific thiol group (S2) of heavy meromyosin.
As previously reported when a specific thiol group, S2, of myosin reacts with N-ethylmaleimide (NEM), its Ca2+-ATPase activity is decreased. Therefore, the reactivity of S2 can be estimated by measuring the decrement of the enzymatic activity. Using the change in the reactivity as a structural probe, we investigated whether F-actin affects the conformation around the region containing S2 under physiological conditions (at neutral pH and low ionic strength). 1. Experiments were carried out with heavy meromyosin (HMM), S1 of which had heen blocked with NEM, to observe the reactivity of S2 alone. In the experiments done in the presence of F-actin, the Ca2+-ATPase activity was measured using the heavy meromyosin fraction after actin had been removed by centrifugation and gel filtration. 2. ATP and other nucleotides activated the reactivity of S2 in the presence of Mg2+. On the other hand, F-actin markedly activated the reactivity of S2 which had been increased by ATP, but not by the other nucleotides. 3. The above cooperative action of F-actin with ATP was not observed in the presence of Ca2+ instead of Mg2+, or above 0.2 M KCl. These results suggest that the S2 region of the myosin molecule is a key region in the molecular interaction of the actin myosin-ATP system under physiological conditions.
D000199:Actins / Q000502:physiology; D000244:Adenosine Diphosphate / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D004789:Enzyme Activation; D005033:Ethylmaleimide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D015879:Myosin Subfragments / Q000378:metabolism; D009218:Myosins / Q000378:metabolism; D009994:Osmolar Concentration; D011189:Potassium Chloride / Q000494:pharmacology; D011487:Protein Conformation; D011817:Rabbits; D012265:Ribonucleotides / Q000494:pharmacology*
D000199:Actins; D015879:Myosin Subfragments; D012265:Ribonucleotides; D000244:Adenosine Diphosphate; D011189:Potassium Chloride; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D009218:Myosins; D005033:Ethylmaleimide; D002118:Calcium