Actomyosin content of Physarum plasmodia and detection of immunological cross-reactions with myosins from related species.
The content of myosin in plasmodia of the myxomycete Physarum polycephalum was measured by an immunological technique, quantitative microcomplement (C') fixation. Migrating plasmodia (starved after growth on rolled oats) contained 0.60 +/- 0.08 (SD) mg myosin per g fresh plasmodia. Myosin comprised 0.77% +/- 0.05 (SD) of the total plasmodial protein. When total plasmodial proteins were separated by electrophoresis on SDS-polyacrylamide gels, a large amount of protein appeared in a band comigrating with muscle actin. Densitometry performed after Coomassie blue staining indicated that as much as 15-25% of the total protein in the plasmodium could be actin. This gives an actin/myosin ratio by weight in the myxomycete plasmodium as high as 19-33, a very "actin-rich" actomyosin compared with rabbit skeletal muscle actomyosin with an actin/myosin ratio of 0.6. Starvation stimulates rapid migration and is correlated with a higher percent of both myosin and actin in the total protein of the plasmodium compared with normally growing cultures. Immunological cross-reaction of myosins from a variety of species was measured by C' fixation using an antiserum produced against purified native myosin from P. polycephalum. Although myxomycete and vertebrate striated muscle myosins have very similar morphological and biochemical properties, and apparently possess similar binding properties to F-actin, only myosins from myxomycetes in the order Physarales, rather closely related to P. polycephalum, gave detectable cross-reactions. This finding suggests that many amino acid sequences in myosin have been variable during evolution.
D000199:Actins / Q000032:analysis*; D000818:Animals; D000941:Antigens; D002645:Chickens; D003168:Complement Fixation Tests; D003429:Cross Reactions; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D009132:Muscles / Q000032:analysis; D009218:Myosins / Q000032:analysis* / Q000276:immunology; D009235:Myxomycetes / Q000032:analysis*; D010804:Physarum / Q000032:analysis*; D011817:Rabbits; D013045:Species Specificity
D000199:Actins; D000941:Antigens; D009218:Myosins