A method to describe enzyme-catalyzed reactions by combining steady state and time course enzyme kinetic parameters |
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Authors: | Ryan Walsh Earl Martin Sultan Darvesh |
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Institution: | 1. Department of Chemistry, Carleton University, 203 Steacie Building, 1125 Colonel By Drive, Ottawa, Ontario, Canada K1S 5B6;2. Department of Chemistry, Mount Saint Vincent University, Halifax, Nova Scotia, Canada;3. Department of Medicine (Neurology and Geriatric Medicine), Dalhousie University, Halifax, Nova Scotia, Canada;4. Department of Anatomy & Neurobiology, Dalhousie University, Halifax, Nova Scotia, Canada;5. Department of Chemistry, Dalhousie University, Halifax, Nova Scotia, Canada |
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Abstract: | BackgroundComplete analysis of single substrate enzyme-catalyzed reactions has required a separate use of two distinct approaches. Steady state approximations are employed to obtain substrate affinity and initial velocity information. Alternatively, first order exponential decay models permit simulation of the time course data for the reactions. Attempts to use integrals of steady state equations to describe reaction time courses have so far met with little success.MethodsHere we use equations based on steady state approximations to directly model time course plots.ResultsTesting these expressions with the enzyme β-galactosidase, which adheres to classical Michaelis–Menten kinetics, produced a good fit between observed and calculated values.General significanceThis study indicates that, in addition to providing information on initial kinetic parameters, steady state approximations can be employed to directly model time course kinetics.Integrated forms of the Michaelis–Menten equation have previously been reported in the literature. Here we describe a method to directly apply steady state approximations to time course analysis for predicting product formation and simultaneously obtain multiple kinetic parameters. |
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Keywords: | β-galactosidase Enzyme kinetics Equation modeling Global data fitting |
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