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Bryant Miles
Reversible versus Irreversible Inhibitiors.
There are many types of enzyme inhibitors. Inhibitors interact with enzymes in one of
two ways, reversibly and irreversibly. Reversible inhibitors interact with the enzyme
through noncovalent interactions and associate and dissociate with the enzyme.
Irreversible inhibitors form stable covalent bonds with the enzyme which block the active
site from binding substrate. The consequence of irreversible enzyme inhibition is a
decrease in the concentration of enzyme active sites.
I. Reversible Inhibitors.
Three categories of reversible enzyme inhibitors are competitive, noncompetitive and
mixed inhibitors.
Competitive inhibitors compete with the substrate for the same binding site on the
enzyme. The inhibitor is prevented from binding to the active site if the active site is
already occupied by a substrate molecule. Thus, increasing the substrate concentration
favors the binding of substrate to the enzyme over the inhibitor, I.
E + S
+
I
Ki
EI
k1
k2
ES
k3
E +P
Ki =
[ E ][ I ]
[ E ][ I ]
; [ EI ]K i = [ E ][ I ];[ EI ] =
Ki
[ EI ]
d [ E ] d [ ES ] d [ EI ]
=
=
=0
dt
dt
dt
d [ ES ]
= k1[ E ][ S ] k 2 [ ES ] k 3 [ ES ] = 0
dt
k1[ E ][ S ] = k 2 [ ES ] + k 3 [ ES ]
[E] =
(k 2 + k 3 )
[ ES ]
k1[ S ]
[ E0 ] = [ E ] + [ ES ] + [ EI ]
[ E0 ] = [ E ] + [ ES ] +
[ E0 ] =
[ E ][ I ]
Ki
[ I ](k 2 + k 3 )
(k 2 + k 3 )
[ ES ]
[ ES ] + [ ES ] +
k1[ S ]
K i k1[ S ]
(k + k 3 ) [ I ](k 2 + k 3 )
[ ES ]
[ E0 ] = 1 + 2
+
k
S
K
k
S
[
]
[
]
i 1
1
K k [ S ] + K i (k 2 + k 3 ) + [ I ](k 2 + k 3 )
[ ES ]
[ E0 ] = i 1
K i k1[ S ]
K i k1[ S ][ E0 ]
= [ ES ]
K i k1[ S ] + K i (k 2 + k 3 ) + [ I ](k 2 + k 3 )
[ ES ] =
v=
[ S ][ E0 ]
(k + k 3 ) [ I ](k 2 + k 3 )
[S ] + 2
+
k1
K i k1
dP
= k 3 [ ES ] =
dt
k 3 [ S ][ E0 ]
(k + k 3 ) [ I ](k 2 + k 3 )
[S ] + 2
+
k1
K i k1
Vmax = k 3 [ E0 ]; Km =
v=
v=
(k 2 + k 3 )
k1
Vmax [ S ]
Vmax [ S ]
=
[ I ]K m
[I ]
[S ] + K m +
[ S ] + K m 1 +
Ki
Ki
Vmax [ S ]
[I ]
[ S ] + K m 1 +
Ki
[I ]
[ S ] + K m 1 +
K i
Vmax [ S ]
K m [I ]
1
1
1 +
v=
; =
=
+
Vmax [ S ]
Vmax Vmax [ S ] K i
[I ] v
[ S ] + K m 1 +
Ki
Noncompetitive inhibitors
Noncompetitive is when the binding of the inhibitor has no effect on the binding of
substrate. That is I and S bind to different sites on the enzyme such that the binding of I
has no effect of the binding of S.
Ki'
Ki
E+I
EI
ES + I
ESI
For pure noncompetitive inhibition Ki=Ki.
The characteristic of a noncompetitive
inhibitor is that it decreases the maximum rate
of the reaction without affecting the Km. The
inhibitor inhibits a fraction of the enzyme
regardless of the substrate concentration.
Lineweaver-Burk plots of noncompetitive
inhibitors all intersect the x-axis at the same
point which corresponds to -1/Km.
Shown above are two Lineweaver-Burk plots of mixed noncompetitive inhibition. Note
that both the slopes and the intercepts change in the presence of I. The first curve on the
left is the situation where Ki < Ki. The graph on the right corresponds to Ki >Ki.
Uncompetitive Inhibition
There some cases where the inhibitor will only bind to the enzyme-substrate complex and
not to the free enzyme. These types of inhibitors are called uncompetitive inhibitors.
These inhibitors are common for enzymes that bind multiple substrates.
k1
k3
E + S
ES
E +P
k2
+
I
Ki
ESI
The Lineweaver-Burk Plots of these inhibitors are a series of parallel lines.
An example of a suicide substrate is Penicillin. The cell walls of bacteria are formed by
macromolecules called peptidoglycan which consists of linear polysaccharide chains
(NAG-NAM)x that are cross linked by short peptides. The cross-linking reaction is
catalyzed by an enzyme called glycoprotein peptidase. The enzyme catalyzed crosslinking reaction involves the formation of an acyl enzyme intermediate.
The antibiotic penicillin covalently reacts with an active site serine if glycoprotein
peptidase. Penicillin effectively blocks cell wall synthesis and makes the cells
susceptible to osmotic lysis.
Penicillin is an effective inhibitor of glycoprotein peptidase
because it resembles the transition state of the normal
substrate RDAlaDAla. (A) is the crystal structure of
penicillin. (B) is the proposed transition state of the
transpeptidase reaction.