Cardiac myosin from pig heart ventricle. Purification and enzymatic properties.
A method is described for the preparation of high purity myosin from the left ventricle of pig heart. The purified myosin was free from nucleic acid, actin, tropomyosin, troponin, the 150,000 molecular weight protein and other contaminants. Analyses of subunits in the purified myosin were carried out on 3.5% acrylamide gel with 0.1% SDS. Of the total protein present in myosin, 11.3% was in the light chains; light chain 1 (LC1), 5.9% and light chain 2 (LC2), 5.4%. Urea gel electrophoresis of the purified myosin showed three closely spaced bands corresponding to the 20,000 dalton, the charge-modified 20,000 dalton and the phosphorylated 20,000 dalton components. The properties of the Ca2+-activated and K+-activated ATPases [EC 3.6.1.3] of the purified myosin were also studied. The Km values were 27 and 55 muM and the Vmax values were 0.263 and 0.317 mumole P1/mg/min for the Ca2+-activated and K+-activated ATPases, respectively. The pH-activity profiles and the effects of SH modification were of the skeletal myosin type except that the activities were lower.
D000251:Adenosine Triphosphatases / Q000032:analysis*; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002729:Chloromercuribenzoates / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D009206:Myocardium / Q000032:analysis*; D009218:Myosins / Q000302:isolation & purification*; D011188:Potassium / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013552:Swine
D002729:Chloromercuribenzoates; D013438:Sulfhydryl Compounds; D000251:Adenosine Triphosphatases; D009218:Myosins; D011188:Potassium; D002118:Calcium