enzyme_kinetics

enzyme_kinetics - Enzymes: Catalysis and Kinetics Why Study...

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Unformatted text preview: Enzymes: Catalysis and Kinetics Why Study Enzyme Kinetics? Quantitative description of biocatalysis Determine the order of binding of substrates Elucidate acid-base catalysis Understand catalytic mechanism Find effective inhibitors Understand regulation of activity Kinetic Concepts For the reaction A + B P Rate = - [ ] = - [B] = [P] t t t Rate = k [A] [B] For the reaction aA + bB P Rate = k [A] a [B] b What assumption are we making? Order of the reaction: sum of the exponents Rate = k [A] Rate = k [A] [B] Rate = k [A] o = k Effect of pH and temperature enzymes have a temperature optimum enzymes have a pH optimum what factors influence the shape and position of this curve? The overall stability of the enzyme, the environment in which the enzyme functions and the pKa of any ionizable group participating in catalysis (a) Tells you (b) tells you Enzyme assays All the following discussion require a method to actually measure the rates of a reaction. 1. Coloured substrates or products, measure appearance or disappearance of colour, UV absorbance. 2. Coupled assays where product of reaction of interest is coupled to a reaction that produces a coloured product. 3. Incorporation of radioactive label. 4. You did an experiment in lab with a coloured substrate analog for the serine proteases. For enzyme assays and kinetics experiments one measures the initial rate of the reaction when the rate depends only on the substrate concentration Michaelis-Menten Kinetics Assume the reaction S P If the reaction is enzyme catalyzed: E + S ES E + P k 1 k-1 k 2 Michaelis-Menten Kinetics E + S ES E + P What is the rate determinig step?...
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enzyme_kinetics - Enzymes: Catalysis and Kinetics Why Study...

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