A simple method for the isolation of actin from myxomycete plasmodia.
A new, simple method for the isolation of actin from myxomycete plasmodia has been developed. Plasmodium myosin B was incubated at 55 degrees C for 15 min in the presence of ATP or was treated with 90% acetone. By this treatment myosin was denatured completely. Actin was then extracted with a dilute ATP and cysteine solution from the heat- or acetone-treated myosin B. The method is simple and almost pure actin was obtained in high yield. The purified G-actin polymerized to F-actin on addition of 0.1 M KCl or 2 mM MgCl2. The viscosity of the purified F-actin was 8-10 dl/g. The F-actin activated muscle myosin ATPase, and actomyosin synthesized from the F-actin and muscle myosin showed superprecipitation on addition of ATP.
D000096:Acetone; D000199:Actins / Q000302:isolation & purification* / Q000494:pharmacology; D000205:Actomyosin; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate; D000818:Animals; D006358:Hot Temperature; D008722:Methods; D009132:Muscles; D009218:Myosins / Q000302:isolation & purification; D009235:Myxomycetes; D010804:Physarum; D011487:Protein Conformation; D011489:Protein Denaturation; D011817:Rabbits
D000199:Actins; D000096:Acetone; D000255:Adenosine Triphosphate; D000205:Actomyosin; D000251:Adenosine Triphosphatases; D009218:Myosins