Myosin from arterial smooth muscle: isolation following actin depolymerization.
The contractile proteins from arterial smooth muscle are highly soluble, and can be extracted at I = 0.05. However, they can be precipitated by a prolonged dialysis at pH 6 to give an actomyosin with a high, although variable, actin:myosin ratio. The sedimentation behavior of this actomyosin at high ionic strength was examined as a function of pH, protein concentration and composition by preparative ultracentrifugation. Comparisons with synthetic skeletal muscle actomyosins of similar composition demonstrated significant differences in the behaviors of these two systems. It was found that much smooth muscle actomyosin is not dissociated by normally relaxing conditions, and that it sediments at a slower rate than F-actin. The solubility of the supernatant protein (a myosin-enriched actomyosin) in 0.2 M K Cl (pH 7) depended on the pH during centrifugation. A lower solubility was associated only with a higher actin concentration in the supernatant, suggesting a dependence on actin repolymerization. Pure myosin was selectively precipitated from the supernatant by polyethylene glycol-6000, but only when the protein was soluble at low ionic strength. The solubility of purified myosin was similar to that of myosin from striated muscles. A relationship between the presence of depolymerized actin and the high solubility of smooth muscle contractile proteins is suggested.
D000199:Actins / Q000302:isolation & purification; D000205:Actomyosin / Q000302:isolation & purification; D000251:Adenosine Triphosphatases / Q000032:analysis; D000255:Adenosine Triphosphate; D000818:Animals; D002339:Carotid Arteries; D002415:Cats; D011232:Chemical Precipitation; D003956:Dialysis; D004285:Dogs; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009130:Muscle, Smooth / Q000032:analysis*; D009218:Myosins / Q000302:isolation & purification*; D009994:Osmolar Concentration; D011092:Polyethylene Glycols; D011189:Potassium Chloride; D012995:Solubility; D013552:Swine
D000199:Actins; D011092:Polyethylene Glycols; D011189:Potassium Chloride; D000255:Adenosine Triphosphate; D000205:Actomyosin; D000251:Adenosine Triphosphatases; D009218:Myosins