Filament formation by purified Physarum myosin.
Physarum myosin can be separated from actomyosin by ultracentrifugation, and purified by gel filtration. Unlike actomyosin, myosin is soluble in 0.05 M KCl in the pH range of 6-7. However, in the absence of actin, the slime mold myosin can be precipitated in 0.05 M KCl by the addition of millimolar concentrations of CaCl(2). The precipitates consist of aggregated, short bipolar filaments. Magnesium has a similar effect, but results in the precipitation of more loosely packed aggregates. The length of the compact filaments is 0.45 mum; thus, predominantly tail-to-tail, but also some head-to-tail, interactions occur under these conditions. Since the size and shape of these thick filaments are close to those seen in fixed and sectioned ameboid cells and in platelets, all of these filaments are probably composed of myosins.
D000205:Actomyosin / Q000302:isolation & purification; D000251:Adenosine Triphosphatases / Q000302:isolation & purification*; D002122:Calcium Chloride; D002247:Carbon Isotopes; D011232:Chemical Precipitation; D002845:Chromatography; D002850:Chromatography, Gel; D003956:Dialysis; D006863:Hydrogen-Ion Concentration; D008239:Lysine / Q000378:metabolism; D008274:Magnesium; D008722:Methods; D008854:Microscopy, Electron; D009124:Muscle Proteins / Q000302:isolation & purification*; D009218:Myosins / Q000302:isolation & purification; D009235:Myxomycetes / Q000254:growth & development; D012995:Solubility; D014461:Ultracentrifugation
D002247:Carbon Isotopes; D009124:Muscle Proteins; D000205:Actomyosin; D000251:Adenosine Triphosphatases; D009218:Myosins; D008274:Magnesium; D008239:Lysine; D002122:Calcium Chloride