Analysis of negative cooperativity for glutamate dehydrogenase

Abstract

The empirical equation, which describes negative cooperativity in the enzyme kinetics, has been proposed. The equation is obtained from the Michaelis–Menten equation where the Michaelis constant is replaced by the effective Michaelis constant, which is a linear function of the v/ V max ratio ( v is the rate of the enzymatic reaction and V max is the limiting value of v at saturating concentrations of substrate). The equation allows the limiting values of the Michaelis constant at v/ V max → 0 and v/ V max → 1 to be estimated, K 0 and K lim, respectively. The K lim/ K 0 ratio is considered as a quantitative characteristic of negative cooperativity. The applicability of the equation has been demonstrated for the kinetic data obtained for glutamate dehydrogenases from various sources (negative kinetic cooperativity for coenzyme). The negative cooperativity for the functions of saturation of protein by ligand is also analyzed. The data on binding of spin-labeled NAD, NADH, and NADPH by beef liver glutamate dehydrogenase are used as examples.