Abstract
Most enzyme kinetic experiments are carried out under pseudo-first-order conditions, that is, when one of the reactant species (the enzyme or the substrate) is in a large excess of the other species. More accurate kinetic information about the system can be gained without the restrictions of the pseudo-first-order conditions. We present a practical and general method of analysis of the common two-step rapid equilibrium Michaelis-Menten mechanism. The formalism is exact in that it does not involve any other approximations such as the steady-state, limitations on the reactant concentrations or on reaction times. We apply this method to the global analysis of kinetic progress curves for bovine alkaline phosphatase assays carried out under both pseudo-first-order and pseudo-second-order conditions.
Original language | English |
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Pages (from-to) | 276-279 |
Number of pages | 4 |
Journal | Analytical Biochemistry |
Volume | 387 |
Issue number | 2 |
DOIs | |
State | Published - Apr 15 2009 |
Keywords
- Alkaline phosphatase
- Kinetic analysis
- Michaelis-Menten
- Progress curve
- Rapid equilibrium
- Rate law
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology