9 - Enzyme Kinetics.pdf - Principles of Enzyme Catalysis I...

Info icon This preview shows pages 1–4. Sign up to view the full content.

View Full Document Right Arrow Icon
3/7/2017 1 1 Principles of Enzyme Catalysis Principles of Enzyme Catalysis I) Enzyme Kinetics II) Inhibition of Enzyme Activity III) Second-Order Reactions IV) Mechanisms of Enzyme Action V) Example: Serine Proteases VI) Enzyme Regulation 2 I) Enzyme Kinetics Michaelis-Menten equation can be used as long as: [E] is constant Formation of EA complex. Breakdown of EA to form products is assumed to be slower than ( i ) formation of EA and ( ii ) breakdown of EA to re-form E and A. V max = k 3 [E t ] Steady-state: concentration of EA complex is constant ( assumption 1 ) Only the initial rate V 0 is measured ( k 4 can be ignored, no inhibition or reversibility - assumption 2 ) [A] >> [E] and A concentration is considered constant ( assumption 3 ) 𝑉 0 = 𝑉 𝑚𝑎𝑥 [?] 𝐾 𝑚 + [?] 𝐸 + ? 𝑘 1 𝑘 2 𝐸? 𝑘 3 𝑘 4 𝐸 + ? 3 1) Mix enzyme and substrate and follow the reaction 2) Determine V 0 at different substrate concentrations 3) Make plots of V 0 vs. [A] or 1/V 0 vs. 1/[A] Enzyme Assay 4
Image of page 1

Info icon This preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon