Professional Documents
Culture Documents
Campbell
Shawn O. Farrell
international.cengage.com/
Chapter Six
The Behavior of Proteins: Enzymes
H2 O 2 H2 O + O2
Temperature dependence of catalysis
• Temperature can also
catalyze reaction (increase
rate)
E + S ES
enzyme-substrate
complex
Binding Models
• Two models have been developed to describe
formation of the enzyme-substrate complex
• Or alternatively [ES] =
[E] T [S]
KM + [S]
Michaelis-Menten Model (Cont’d)
• In the initial stages, formation of product depends only on the
rate of breakdown of ES
k 2[E]T [S]
Vinit = k 2 [ES] =
KM + [S]
• If substrate concentration is so large that the enzyme is
saturated with substrate [ES] = [E]T
Vmax [S]
V= (an equation for a hyperbola)
KM + [S]
• Can be transformed into the equation for a straight line by taking
the reciprocal of each side
1 = KM + [S] KM [S]
= +
V Vmax [S] Vmax [S] Vmax [S]
1 = KM 1
+
V Vmax [S] Vmax
Lineweaver-Burk Plot
• The Lineweaver-Burke plot has the form y = mx + b, and is the
formula for a straight line
1 KM 1 1
= • +
V Vmax [S] Vmax
y = m • x + b
• a plot of 1/V versus 1/[S] will give a straight line with slope of
KM/Vmax and y intercept of 1/Vmax
• such a plot is known as a Lineweaver-Burk double reciprocal
plot
Lineweaver-Burk Plot (Cont’d)
• KM is the dissociation constant for ES; the greater the value of
KM, the less tightly S is bound to E
EI I + E + S ES P
[E][I]
EI I + E KI =
[EI]
Competitive Inhibition
No inhibition
1 = KM • 1 + 1
V Vmax S Vmax
y = m • x + b
y = m • x + b
+S
E ES E + P
-S
-I +I -I +I
+S
EI ESI
-S
Vmax
I
V max =
1 + [I]/K I
A Lineweaver-Burke Plot for
Noncompetitive Inhibition
• Because the inhibitor does not interfere with binding of substrate
to the active site, KM is unchanged
• Increasing substrate concentration cannot overcome
noncompetitive inhibition
No inhibition
1 = KM • 1 + 1
V Vmax S Vmax
y = m• x + b
In the presence of a noncompetitive inhibitor
1 = KM [I] 1 1 [I]
1 + + 1 +
V Vmax KI S Vmax KI
y = m • x + b
A Lineweaver-Burke Plot for
Noncompetitive Inhibition (Cont’d)
Other Types of Inhibition
• Uncompetitive- inhibitor can bind to the ES complex
but not to free E. Vmax decreases and KM decreases.