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Determination of the rapid-equilibrium kinetic parameters for the enzyme-catalyzed reaction A + mB -> products with the minimum number of velocity mesurements

Robert A. Alberty
Organization: Massachusetts Institute of Technology
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This package shows how to estimate kinetic parameters for two types of enzyme-catalyzed reactions: A + 2B -> products and A + 3B -> products using the minimum number of velocity measurements. Both ordered and random mechanisms are considered. The expressions for the velocities of these reactions are derived using Solve. Eight rapid-equilibrium rate equations are derived. Eight programs, also involving Solve, are provided to calculate kinetic parameters from measured velocities for eight different mechanisms. These programs are also used to calculate effects of 5% errors measured velocities and substrate concentrations. These effects are summarized in tables. The choices of pairs of substrate concentrations are guided by 3D plots of velocities versus [A] and [B], and 3D plots of partial derivatives versus [A] and [B]. When a reaction is studied for the first time there is an advantage in using a random program because it can yield the kinetic parameters for an ordered mechanism. For random mechanisms, thermodynamics makes it possible to calculate five kinetic parameters from four velocities because of thermodynamics. These calculations can be extended to reactions involving more B molecules and to mechanisms involving step-wise binding of B molecules.

*Science > Biochemistry

biochemistry, enzyme-catalyzed reactions, kinetics, kinetic parameters, enzyme mechanisms
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KinAB234-30-08.nb (2.4 MB) - Mathematica Notebook