Application of the Van Slyke-Cullen irreversible mechanism in the analysis of enzymatic progress curves.
For enzymatic progress curves conforming to the Michaelis-Menten mechanism E+Sright harpoon over left harpoonES-->E+P, the minimal fitting model cast as a system of numerically integrated differential equations is the simplified, irreversible Van Slyke-Cullen mechanism E+S-->ES-->E+P. The best-fit value of the bimolecular association rate constant is identical to the specificity constant kcat/KM. An illustrative example involves a fluorogenic continuous assay of the HIV protease, analyzed by the differential-equation oriented software package DYNAFIT [P. Kuzmic, Anal. Biochem. 237 (1996) 260].
Kuzmic
Petr
P
BioKin Ltd., 15 Main Street Suite 232, Watertown, MA 02472, USA. pksci01@biokin.com
D019295:Computational Biology; D003198:Computer Simulation; D016333:HIV Protease / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D011485:Protein Binding; D012984:Software; D013379:Substrate Specificity
D016333:HIV Protease