In vitro stability and protein composition of thick filaments from insect flight muscles

Abstract

Thick and thin filaments of synchronous and asynchronous insect flight muscles were separated by density gradient centrifugation. A good release of myofilaments from myofibrils was obtained by sonication of myofibrils in relaxing solution with pH 6.1 (locust), pH 6.4 (honeybee) and pH 6.6 (fleshfly), respectively. Thick filaments but not thin filaments were dissolved, if sucrose gradient centrifugation was used to separate the filaments. Thus, sucrose gradients are the medium of choice if actin filaments are to be purified. Glycerol-containing gradients selectively dissolved myosin filaments from fleshfly muscles. The stability of the myosin filaments of all muscles was sufficient in gradients with 10–30% formamide.