‫ﺑﺴﻢ ﷲ اﻟﺮﺣﻤﻦ اﻟﺮﺣﯿﻢ‬

Enzyme Inhibition

Bioc. 231

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 Enzyme Inhibition
• One of the ways of altering enzyme activity is through
compounds binding in the active site. If these compounds
are not part of the normal reaction, they inhibit the
enzyme.
§ Inhibitors are compounds that decrease the rate of an
enzyme-catalyzed reaction by binding to the enzyme.
• The study of enzyme inhibition has enhanced our
knowledge of specificity and the nature of functional
groups at the active site.
• The activity of certain enzymes is regulated by a feedback
mechanism such that an end product inhibits the enzyme’s
function in an initial stage of a sequence of reactions

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 Enzyme Inhibition
§ The action of an inhibitor on an enzyme can be described as either
irreversible or reversible.
§ In irreversible inhibitions, inhibition progressively increases with time.
• Complete inhibition results if the concentration of the irreversible
inhibitor exceeds that of the enzyme.
• Irreversible inhibitors bind to enzymes through covalent bonds.
§ In reversible inhibition, an equilibrium exists between the enzyme and
the inhibitor.
• Reversible inhibitors typically bind to enzymes through non covalent
bonds, thus dilution of the enzyme–inhibitor complex results in
dissociation of the reversibly bound inhibitor, and recovery of enzyme
activity.
• Reversible inhibitors are generally classified as competitive, non-
competitive, or uncompetitive with respect to their relationship to a
substrate of the enzyme.
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 Reversible Inhibition:
1. Competitive Inhibition
• A competitive inhibitor
competes with a substrate for
binding at the same active site
and therefore is usually a close
structural analog of the
substrate.
• Note that the complex EI does
not react with S to form
products.

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 Reversible Inhibition:
1. Competitive Inhibition

• The dissociation constant for the reaction between E and I is KI,

where

• Applying the steady-state approximation for ES, we obtain

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 Reversible Inhibition:
1. Competitive Inhibition
• The Lineweaver–Burk
equation is given by

• The slope is enhanced by

the factor (1 + [I]/KI). The
intercept on the 1/v0 axis
is the same because Vmax
does not change.

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 Reversible Inhibition:
1. Competitive Inhibition
• A well-known example of a competitive inhibitor is malonic acid,
CH2(COOH)2, which competes with succinic acid in the dehydrogenation
reaction catalyzed by succinic dehydrogenase.
• Because malonic acid resembles succinic acid in structure, it can combine
with the enzyme, although no product is formed in this reaction.

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 Reversible Inhibition:
1. Competitive Inhibition
• To overcome competitive inhibition, we need
to increase the substrate concentration;
when the substrate concentration is increased
to sufficiently high level, the substrate-binding
sites are occupied by substrate and inhibitor
molecules cannot bind.

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 Reversible Inhibition:
1. Competitive Inhibition
• Competitive inhibitors increase the apparent KM of the
enzyme because they raise the concentration of substrate
necessary to saturate the enzyme. They have no effect on
Vmax.

A. Effect of a competitive inhibitor on the reaction velocity (v0) versus substrate ([S]) plot.
B. Lineweaver-Burk plot of competitive inhibition of an enzyme. 9
 Reversible Inhibition:
1. Competitive Inhibition
• The inhibited and uninhibited reactions show different x-axis
intercepts, indicating that the apparent KM is increased in the
presence of the competitive inhibitor because -1/ KM moves
closer to zero from a negative value.

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 Reversible Inhibition:
2. Non Competitive Inhibition
• A noncompetitive inhibitor
binds an enzyme at a site
other than the active site;
therefore, it can bind to both
the free enzyme and the
enzyme–substrate complex .
• The binding of the inhibitor
has no effect on the substrate
binding, and vice versa.

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 Reversible Inhibition:
2. Non Competitive Inhibition
• Non competitive inhibition
also may be irreversible if the
inhibitor destroys part of the
enzyme involved in catalytic
activity.
• Neither EI nor ESI forms
products.
• Because I does not interfere
with the formation of ES,
noncompetitive inhibition
cannot be reversed by
increasing the substrate
concentration.
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 Reversible Inhibition:
2. Non Competitive Inhibition
• The initial rate is given by:

• Vmax has been reduced by the factor (1 + [I]/KI)

but KM is unchanged.

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 Reversible Inhibition:
2. Non Competitive Inhibition
• The Lineweaver–Burk
equation becomes

• The plot of 1/v0 versus

1/[S] gives a straight
line with an increase in
slope and intercept on
the 1/ v0 axis.

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 Reversible Inhibition:
2. Non Competitive Inhibition
• The binding of the heavy metal (e.g. Hg, Ag, Pb)
shows noncompetitive inhibition.

• Ferrochelatase, an enzyme that catalyzes the

insertion of Fe2+ into protoporphyrin (a precursor
of heme), is an example of an enzyme sensitive to
inhibition by lead (Pb). Lead forms covalent
bonds with the sulfhydryl side chains of cysteine
in proteins.
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 Reversible Inhibition:
2. Non Competitive Inhibition
• Noncompetitive inhibitors decrease the apparent Vmax of the
reaction.
• Noncompetitive inhibitors do not interfere with the binding of
substrate to enzyme. Thus, the enzyme shows the same Km in the
presence or absence of the noncompetitive inhibitor.

A. Effect of a noncompetitive inhibitor on the reaction velocity (vo) versus substrate ([S]) plot.
B. Lineweaver-Burk plot of noncompetitive inhibition of an enzyme. 17
 Reversible Inhibition:
3. Uncompetitive Inhibition
• An uncompetitive inhibitor
does not bind to the free
enzyme; instead, it binds
reversibly to the enzyme–
substrate complex to yield an
inactive ESI complex.
• Increasing substrate
concentration results in more
ES complexes to which the
inhibitor binds and, thereby,
increases the inhibitions.

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 Reversible Inhibition:
3. Uncompetitive Inhibition
• The ESI complex does not
yield a product.
• Because I does not interfere
with the formation of ES,
uncompetitive inhibition
cannot be reversed by
increasing the substrate
concentration.

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 Reversible Inhibition:
3. Uncompetitive Inhibition
• The initial rate is given by:

• Both Vmax and KM have been reduced by the

factor (1 + [I]/KI).

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 Reversible Inhibition:
3. Uncompetitive Inhibition
• The Lineweaver–Burk
equation is given by:

• A straight line is obtained

by plotting 1/v0 versus
1/[S] at constant [I].
• The intercept on the 1/v0
axis is altered by the
factor (1 + [I]/KI), but the
slope remains the same.
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 Reversible Inhibition:
3. Uncompetitive Inhibition
• Uncompetitive inhibitors decrease the Vmax and Km values of
the reaction.

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