Received: February 26, 2007

In Final Form: March 30, 2007

Abstract:

Equations obtained in part I for the free-energy barrier to one-step enzymatic reactions between bound reactants are discussed. The rate is expressed in terms of _o (protein reorganization energy), G (standard free energy of reaction of the H-transfer step), bond breaking/bond forming term, w (work terms), and H-transmission property. Two alternative approximations for the coupling of the bond breaking/bond forming and protein are distinguished experimentally in favorable cases by the G where the maximum deuterium kinetic isotope effect occurs. Plots of log rate versus G and properties such as S* and S are discussed. The weak or zero T-dependence of the kinetic isotope effect for wild-type enzymes operating under physiological conditions is interpreted in terms of vanishing (or isotopically insensitive) w plus transfer from the lowest H-state. Static and dynamic protein flexibility is discussed. While the many correlations accessible for electron transfers are not available for H-transfers in enzymes, a combination of experiment, computation, and analytical approaches can assist in evaluating the utility of the present equations and in suggesting further experiments and computations. A protein reorganization energy _o is obtained in the literature from the extended valence bond formalism where diabatic electronic states are used. A method is suggested for extracting it when instead a bond distance difference coordinate is used. The results may provide a bridge between the two approaches.

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