Enzyme kinetic evidence of active-site involvement in the interaction between aldolase and muscle myofibrils.

The interaction of aldolase with the myofibrillar matrix of rabbit skeletal muscle has been investigated by means of its effect on kinetic parameters for the enzyme-catalyzed cleavage of fructose 1,6-bisphosphate. Involvement of the active site in the enzymic interaction with the thin filament of muscle is indicated, the association constant for competitive inhibition of catalysis (420,000 M-1) being in excellent agreement with the value of 410,000 M-1 obtained under the same conditions (pH 6.8, I 0.16) from partition equilibrium studies of the aldolase-myofibril interaction (Kuter, M.R., Masters, C.J. and Winzor, D.J. (1983) Arch. Biochem. Biophys. 225, 384-389). A second kinetic study, designed to take into greater account the inhibitory effects of substrate and other phosphate-containing metabolites on the interaction of enzyme with myofibrils, has substantiated further the concept of aldolase existing as an equilibrium mixture of cytoplasmic and filament-bound forms in muscle tissue.