Properties of Physarum myosin purified by a potassium iodide procedure.
Myosin has been purified free of actin from Physarum actomyosin by a two step adaptation of the classical potassium iodide method for depolymerizing actin. On 12% sodium dodecyl sulfate (SDS) gels, the single major slowly moving protein band present in the calcium activated adenosine triphosphatase peak (90% pure) is associated with two fast moving bands of molecular weights of approximately 17,000 and 21,000 daltons, respectively. Densitometry shows the molar ratio of heavy chains to the 21,000 and 17,000 dalton chains on the gels to be 1:2:1. The highly purified myosin forms filaments up to 2.5 microm long in the presence of 5 mM magnesium and 0.05 M KCl. Calcium ions were not required for the formation of long filaments from this highly purified myosin. At low ionic strength (0.05 M KCl) the magnesium ATPase of the highly purified myosin is activated four- to tenfold by muscle actin. The extent of activation is a function of the actin concentration and levels off at high levels of actin. In 0.1 mM calcium salts the ATPase activity is approximately 60% of that in 1 mM EGTA. In summary, Physarum myosin is similar to a number of muscle myosins as well as to platelet and fibroblast myosin, which all possess light chains of two different molecular weights associated with the heavy chains. Under ionic conditions close to those in vivo, highly purified Physarum myosin aggregates into long filaments.
D000199:Actins; D000251:Adenosine Triphosphatases / Q000032:analysis / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002614:Chelating Agents; D002850:Chromatography, Gel; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005069:Evaluation Studies as Topic; D007454:Iodides; D007700:Kinetics; D008274:Magnesium; D008722:Methods; D008854:Microscopy, Electron; D008970:Molecular Weight; D009132:Muscles; D009218:Myosins / Q000032:analysis* / Q000302:isolation & purification; D009235:Myxomycetes / Q000032:analysis* / Q000201:enzymology; D011188:Potassium; D011485:Protein Binding; D012967:Sodium Dodecyl Sulfate; D013045:Species Specificity; D013997:Time Factors
D000199:Actins; D002614:Chelating Agents; D007454:Iodides; D012967:Sodium Dodecyl Sulfate; D000251:Adenosine Triphosphatases; D009218:Myosins; D008274:Magnesium; D011188:Potassium; D002118:Calcium