Isolation of F-actin filaments. Comparison of F-actin filament preparations from normal and dystrophic mouse muscle.
To investigate the dystrophic influence on the characteristics of actin, a method for the isolation of F-actin filaments from the skeletal muscle of small sizes, i.e., less than 0.5 g, was devised. In this method, minced muscle was treated with collagenase and hyaluronidase, and the isolated filaments were washed with adenosine triphosphate (ATP). Upon examination in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the ATP-washed filaments showed a protein component identical in mobility to actin in untreated myofibrils or to that prepared by the conventional method. Electron microscopic appearances of the filaments were similar to those of F-actin filaments described in the literature. The dimensions of the filaments were 0.5--2.5 micrometer in length and 60--70 A in diameter. The ability to activate the Mg-adenosine triphosphatase or myosin was found to be Ca2+ independent. In all aspects of the above characteristics, the filaments from leg muscles of 129/Re dydy dystrophic mice and their litter mates were observed to be identical.
D000199:Actins / Q000302:isolation & purification*; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D005260:Female; D007866:Leg; D008297:Male; D051379:Mice; D009132:Muscles / Q000032:analysis* / Q000201:enzymology; D009137:Muscular Dystrophy, Animal / Q000378:metabolism*
D000199:Actins; D000251:Adenosine Triphosphatases