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Enzyme Kinetics

Learning Goals:
1. The rate of an enzyme catalyzed reaction can be described by the Michaelis-Menton equation (know
2. Enzyme catalyzed reactions are characterized by two key constants: Vmax and Km (each substrate has a Km) 
Lineweaver-Burk plots
3. Inhibitors of enzymes (drugs): basis for competitive/noncompetitive, allosteric inhibitors or inactivators.
I. The BIG Picture
a. Michaelis Menton equation: vo = Vmax[S]/([S]+Km)
b. Vmax is proportional to E (adding E  increased Vmax  increased observed velocity)
c. As [S] gets large  vo=Vmax
d. Inhibitors (drugs) mimic substrate, bind to binding site (competitive, uncompetitive, non-competitive,
allosteric inhibitors, inactivators)
II. Activation Energy: Enzymes lower activation energy
III. Kinetics of Ordinary Chemical Reactions: A+B  P+Q
a. Law of mass action: observed rate is proportional to [A][B]  vo=K[A][B]
b. Reaction order: elementary reaction- equation describes what really happens, reaction order equals the number
of species that collide for reaction to occur
IV. Kinetics of Enzyme-Catalyzed reactions
a. Two steps: substrate binds enzyme, complex of ES undergoes reaction to form P
b. Simplifying assumptions
i. Initial Rate Assumption: [S] = amount of S added (none is used up before measurement), no P  no
reversible reaction. ES  E+P is irreversible.
ii. Steady State Assumption: [ES] is constant during the time P is being formed
iii. Derivation of the Michaelis-Menton equation
c. Implications of Michaelis-Menton equation
i. Increase catalytic rate constant or total enzyme  vo increases
ii. [S] << Km  vo=Vmax[S]/Km  linearly proportional to [S]
iii. [S] >> Km  vo=Vmax  rate is independent of [S] (enzyme is saturated)
iv. Km = [S] where vo=1/2Vmax
v. Dissociation constant KD is [products]/[reactants] = k-1/k1; if chemistry is slow relative to dissociation
of S from ES, then KD ~ Km
V. The Lineweaver-Burke Equation
a. Rearranged Michaelis-Menton to give a linear plot
b. Y intercept = 1/Vmax and X intercept = -1/Km
VI. Log-Dose Response curves
a. Inflection point = 1/2Vmax
b. 10x Km concentration  90% Vmax
c. 1/10 Km concentration  10% Vmax
VII. Reversible Inhibitors
a. Competitive inhibitors
i. Binds only to E, not ES
ii. Increases the apparent Km (pulls the equilibrium backwards making it look as if S has a lesser affinity
for E)
iii. As [S] becomes large, I becomes less effective (competitive)  Vmax unaffected
b. Noncompetitive inhibitors
i. I binds to E and ES
ii. Vmax is decreased (essentially decreases the amount of functional enzyme)
iii. High [S] cannot overcome inhibition
c. Uncompetitive inhibitors
i. I binds to ES
ii. Apparent Km is decreased
iii. Vmax is decreased
VIII. Irreversible Inhibitors
a. Bind to their target enzyme irreversibly
b. Inactivator (no reversible binding)
IX. Allosteric Enzymes and Inhibitors
a. An effecter binds to a site other than the active site
b. Cooperativity- do not obey Michaelis-Menton kinetics
c. Multiple interacting subunits