Determination of kinetic parameters of enzyme-catalyzed reactions with a minimum number of velocity measurements

Duggleby [Duggleby, R.G., 1979. Experimental designs for estimating kinetic parameters for enzyme-catalyzed reactions. J. Theor. Biol. 81, 672–684] discussed the “design of several replicate measurements of the velocity at as many experimental conditions as there are parameters to be estimated.” He discussed the application of this method to A→products, without and with competitive inhibition, and commented briefly on A+B→products. The availability of computer applications that can solve large sets of simultaneous equations makes it possible to use this method to calculate kinetic parameters for more complicated enzyme mechanisms. This article is concerned with rapid-equilibrium rate equations, but this method can also be used with steady-state rate equations. Computer programs are provided for the calculation of the three kinetic parameters for ordered A+B→products from three velocity measurements and for the calculation of the four kinetic parameters for random A+B→products from four velocity measurements. Computer programs are also provided for competitive inhibition, uncompetitive inhibition, and mixed inhibition of ordered A+B→products.