9th Class
9th Class
9th Class
10/3/01
The significance of enzyme kinetics Derivatisation of rapid equilibrium equations The steady state assumption and Derivation of the Briggs-Haldane equation Linear transformations and linear plots and their applications Time integrated equation Inhibition kinetics - techniques and uses Nomenclature Rapid kinetics - experimental examples Limits of kinetic rate constants and the rate determining step Useful kinetic shortcuts
References 1. Fersht, Ch. 3. and 4 2. Cleland, Meths Enzymol. 87, 390-405 (1982). pH analysis. The remainder of this volume has considerable useful information for the practicing kineticist. 3. Cleland, adv. Enzymol. 45, 273-387 (1977). Most of what the normal kineticist will ever have to know is to be found here. 4. Segel, "Enzyme Kinetics" (1975), Wiley. Very comprehensive book. 5. King, E.L & Altman C. J. Phys. Chem. 60, 1375-1378 (1956). The King-Altman method for driving kinetic equations. 6. Cleland, Biochemistry 14, 3220 (1975). The net rate constant method for the analysis of linear mechanisms. 7. Cha, J. Biol. Chem. 243, 820-825 (1968). Rapid equilibrium simplification of King Altman procedure. 8. Frieden, Trends in Biochemical Sciences, 4, 181-182.(1994) Numerical integration of rate equations by computer. 9. Northrop, D.B. j. Chem. Ed. 75, 1153-1157 (1998). On the meaning of KM and V/K.
k 2 < < k -1
v=
f ([S])
W h a t a r e t h e r e l a t i o n s b e tK w e n K S? Me and Check Fersht Ch. 3 - A.3.b. (pp 107-108) for opposite a actually more general case.
Competitive inhibition
Graphic presentations
Dixon plot
Non-competitive inhibition
Graphic presentation
Uncompetitive inhibition
Mixed inhibition
EA + B
EAB
kp
E+P
Application
Exp. v
40
V (s-1)
30 20
10 0
2
+
[NAD ] (mM)
0.3
]/V (mM*min.)
0.2
[NAD +
0.1
-1
2
+
[NAD ] (mM)
Quenched flow
Pulse flow