Comparative studies on amino and thiol groups in myosins from different sources.
Myosins from rabbit white and red skeletal, rabbit heart, fish skeletal and chicken gizzard muscles, as well as from human platelets were subjected to trinitrophenylation by trinitrobenzene sulfonate and alkylation by N-ethylmeleimide which affected their amino and thiol groups, respectively. The blocking of amino groups was carried out in the presence or in the absence of Mg-ADP and was followed both spectrophotometrically and enzymatically. Essential amino groups, whose modification throughly changes the enzymic characteristics of myosin, were found in heart and in all skeletal muscle myosins but were absent in myosins from chicken gizzard muscle and from human platelets. The reaction of these amino groups was highly retarded in the presence of Mg-ADP. Alkylation of thiols led to loss of the K+-activated ATPase (ATP phosphohydrolase, EC 3.6.1.3) in all myosins. However, the rate of loss of activity varied from one myosin to another and, for a given myosin, was affected by the presence of nucleotides and by the value of the ionic strength. The change in Ca(2+)-activated ATPase activity (ATP phosphohydrolase, EC 3.6.1.3) on alkylation was influenced by the presence of Mg - ADP during the reaction. In the absence of this nucleotide, the Ca(2+)-ATPase activity increased and reached a plateau as a consequence of modification. The extent of activation largely depended on the origin of the myosin. When alkylation was carried out in the presence of Mg-ADP, the Ca(2+)-ATPase activity as a function of time exhibited a maximum but the descending part of the curve was absent in myosins from heart and gizzard muscles.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000588:Amines / Q000032:analysis; D000818:Animals; D001792:Blood Platelets / Q000201:enzymology; D002118:Calcium / Q000494:pharmacology; D002645:Chickens; D004789:Enzyme Activation / Q000187:drug effects; D005033:Ethylmaleimide / Q000494:pharmacology; D005399:Fishes; D005895:Gizzard, Avian / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D009132:Muscles / Q000201:enzymology; D009218:Myosins / Q000378:metabolism; D009928:Organ Specificity; D011188:Potassium / Q000494:pharmacology; D011817:Rabbits; D013045:Species Specificity; D013438:Sulfhydryl Compounds / Q000032:analysis; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D014302:Trinitrobenzenesulfonic Acid / Q000494:pharmacology
D000588:Amines; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; D014302:Trinitrobenzenesulfonic Acid; D000251:Adenosine Triphosphatases; D009218:Myosins; D005033:Ethylmaleimide; D011188:Potassium; D002118:Calcium