New, rapid methods for purifying alpha-actinin from chicken gizzard and chicken pectoral muscle.
We introduce two new, rapid procedures. One is specifically designed for isolating alpha-actinin from skeletal and the other for isolating alpha-actinin from smooth muscle. Approximately 20 mg of greater than 95% pure alpha-actinin can be obtained/100 g of ground chicken pectoral muscle in just 4 days. The smooth muscle protocol yields 2.7 mg of greater than 99% pure alpha-actinin/100 g of ground gizzard after just 5 days. Differences in protein contaminants and in the extractability of alpha-actinin necessitated the development of separate isolation procedures for the two muscle types. Antibody prepared against the purified gizzard alpha-actinin reacted with alpha-actinin from skeletal, cardiac, and smooth muscle in immunodiffusion. Anti-alpha-actinin reacted only with alpha-actinin from crude extracts of skeletal and smooth muscle on Staph A gels. Anti-alpha-actinin stained Z-bands from skeletal muscle in indirect immunofluorescence microscopy and stress fibers from baby hamster kidney fibroblasts and mouse mammary epithelial cells in the characteristic punctate pattern observed by other workers (Lazarides, E., and Burridge, K. (1975) Cell 6, 289-298). These two methods for purifying alpha-actinin from skeletal and smooth muscle represent a significant improvement over that published previously.
D000185:Actinin / Q000302:isolation & purification*; D000818:Animals; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D005895:Gizzard, Avian / Q000032:analysis*; D006651:Histocytochemistry; D005779:Immunodiffusion; D008970:Molecular Weight; D009124:Muscle Proteins / Q000302:isolation & purification*; D009132:Muscles / Q000032:analysis*; D009928:Organ Specificity
D009124:Muscle Proteins; D000185:Actinin