The sulfhydryl groups involved in the active site of myosin B adenosinetriphosphatase. II. Effect of modification of the Sa thiol group on superprecipitation and clearing.
The effect of Sa modification with NEM, which activates Mg2+-ATPase through an enhancement of the association of actin and myosin, was investigated on the superprecipitation, clearing and Mg2+-ITPase of myosin B with reference to the effect of S1-blocking. 1. Superprecipitation induced by ATP was markedly enhanced by Sa-blocking even at high concentrations of Mg2+ and substrate; this may be due to an increase in the affinity of myosin and actin on blocking Sa. 2. Nevertheless, neither ITP-induced superprecipitation nor Mg2+-ITPase was affected by Sa modification. 3. Blocking of S1 brought about the inhibition of ATP- and ITP-induced superprecipitation and Mg2+-ITPase activity, suggesting that S1-blocking decreases the affinity of myosin and actin. 4. Sa-blocked myosin B showed greater resistance to clearing by ATP, especially in the presence of Ca2+ ions, whereas in the clearing response of actomyosin gel to PPi no difference between Sa-blocked and unmodified myosins B was observed. On the other hand, the clearing response of myosin B became more sensitive to both ATP and PPi on blocking S1. Based on the above results and preliminary data suggesting that Sa is located in LMM, the interaction of myosin filaments and actin filaments under physiological conditions is discussed.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000494:pharmacology; D011232:Chemical Precipitation; D011756:Diphosphates / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D007292:Inosine Nucleotides / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D009218:Myosins / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D011188:Potassium / Q000494:pharmacology; D011817:Rabbits; D013438:Sulfhydryl Compounds / Q000378:metabolism*
D011756:Diphosphates; D007292:Inosine Nucleotides; D013438:Sulfhydryl Compounds; D000255:Adenosine Triphosphate; D010744:Phosphoric Monoester Hydrolases; D000251:Adenosine Triphosphatases; D009218:Myosins; D008274:Magnesium; D005033:Ethylmaleimide; D011188:Potassium; D002118:Calcium