Heat treated myosin does not bind ATPase ‐ inhibiting antibodies

Abstract

Polyclonal antibodies to native chicken pectoral fast-twitch myosin are directed to all subfragments of the molecule ($1, $2 and LMM), as seen in the ELISA and Western blotting techniques. The antibodies inhibit the Ca2+-activated myosin ATPase. Absorption of the antibodies with native myosin abolishes these reactions. Heat treatment of myosin for 2h at 40~ will inactivate myosin ATPase and alter its antibody binding pattern: the binding of antibodies to the rod fractions is reduced, that to the globular head ($1) completely abolished. Thus, these antibodies are useful as sensitive probes for the structural integrity of the myosin head.

Expression of two ATP-gated ion channels, P2X5 and P2X6, in developing chick skeletal muscle

“…The antiserum to chicken myosin (fast twitch glycolytic) (a‐PM) was obtained after injection of the purified immunogen into rabbits. The specificity of this antiserum is described in detail by Gröschel‐Stewart et al (1997). Immunoglobulins G (IgG) were isolated from the immune sera according to the method of Harboe and Ingild (1973).…”

“…Specificity of the P2X antibodies was verified by immunoblotting with chick embryo crude extracts (stages 30–32). (Western blots of the myosin antibodies are shown in Gröschel‐Stewart et al, 1997.) Generation of crude extracts: five embryos between the stages of 30 and 32 were frozen in liquid nitrogen and ground to a fine powder with a mortar and pestle.…”