Last edited: 9/10/2021

1. ENZYME INHIBITION
Enzyme Inhibition: Reversible and Irreversible Inhibition Medical Editor: Jona Frondoso

OUTLINE Two Types of Enzyme Inhibition

I) INTRODUCTION (i) Irreversible

II) TYPES OF REVERSIBLE INHIBITION  Bind to enzymes through covalent bonds [Harvey &
III) IRREVERSIBLE INHIBITION
Ferrier, 2011]
IV) APPENDIX
V) REVIEW QUESTIONS (ii) Reversible
VI) REFERENCES
 Bind to enzymes through non-covalent bonds
[Harvey & Ferrier, 2011]

I) INTRODUCTION  Dilution of the enzyme-inhibitor complex results in

the dissociation of the reversibly bound inhibitor
Enzyme inhibitors have ways of affecting the normal [Harvey & Ferrier, 2011]
process of reaction (Figure 1).

Km and Vmax

Figure 1. Reaction model showing an enzyme reversibly
combining with its substrate to form an ES complex that
subsequently yields product, regenerating the free enzyme.
Where
o E is the enzyme
o S is the substrate
o ES is the enzyme-substrate complex
o P is the product
o k1 is the rate constant for the forward reaction of E
and S forming ES
o k- 1 is the rate constant for the reverse reaction of ES
forming E and S FYI:
o k2 is the rate constant for the forward reaction of ES Km and Affinity
forming E and P
Inhibitor
Any substance that can diminish the velocity of an enzyme-
catalyzed reaction [Harvey & Ferrier, 2011]
(i) Km
Reflects the affinity of the enzyme for a particular substrate
Substrate concentration ([S]) at ½ Vmax
Changes when the equilibrium changes
(ii) Vmax
Maximal velocity
Velocity of the reaction when all enzymes are saturated
by the substrate
Changes when the amount of ES complex formation
changes
(i) Small Km
↑ Affinity of the enzyme for a particular substrate
↓ Concentration is needed to achieve ½ Vmax
(ii) Large Km
↓ Affinity of the enzyme for a particular substrate
↑ Concentration is needed to achieve ½ Vmax

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 II) TYPES OF REVERSIBLE INHIBITION
• Can be competitive, noncompetitive, uncompetitive

(A) COMPETITIVE INHIBITION (B) NONCOMPETITIVE INHIBITION

Competitive inhibitor (CI)
o Structural analog of the actual substrate → can also
bind to the active site
o Forms noncovalent bonds with the active site
 Noncovalent bonds – weak bonds → can still be
broken
o CI bound to the active site → actual substrate can no
longer bind → ↑ Affinity for the CI while ↓ affinity for the
substrate→ ↓ ES complex formation → reaction shifts
to the right (by Le Chatelier’s Principle) → can be
(1) Effect on Vmax
achieved by ↑ substrate concentration (Figure 1)
(1) Effect on Vmax
Normal / Unchanged Vmax
Effect can be reversed by ↑ substrate concentration
↑ Substrate concentration that can bind to the active site
→ overcoming the concentration of competitive inhibitors
→ ↑ ES complex formation → ↑ product formed or the
(2) Effect on Km
amount of products formed when the inhibitor is absent
(Figure 1)
o Means Vmax can still be achieved
(2) Effect on Km
↑ Km
Equilibrium is changed
o Since reaction shifted to the right
More substrate is needed to achieve the same ½ Vmax
Noncompetitive inhibitors (NCI) – inhibitors that bind the
allosteric site of either the free enzyme or the ES
complex
Substrate can still bind to the active site while NCI is bound
to the allosteric site → ESI complex formation → no
products formed
When the inhibitor is released → ES complex formation
→ products formed
↓ Vmax
o Not all enzymes can only be bound to the substrate
o Enzymes can form either ES complex or ESI complex
o Can never be reached Vmax
Some enzymes participate in the ESI complex formation →
↓ ES complex formed →↓ products formed
Normal / Unchanged Km
Because equilibrium for the reaction is not changed or not
thrown off
Reaction can still proceed towards the formation of the
products

Figure 3. Noncompetitive inhibition.

(3) Examples

(i) Monoamine oxidase inhibitors (MAO-I)

Used in depression
Inhibit monoamine oxidase (MAO)
Figure 2. Competitive inhibition.
o MAO – breaks down epinephrine, norepinephrine, and
dopamine
(3) Examples (ii) Oxamic acid
(i) Statins Inhibits lactate dehydrogenase
Lactate dehydrogenase
Used in hypercholesterolemia and hypertriglyceridemia
o Converts lactate to pyruvate
Inhibits hydroxymethylglutaryl Coenzyme A (HMG-
o Pyruvate will eventually be converted to glucose
CoA) synthase

(ii) ACE inhibitors

Used in hypertension
Inhibits angiotensin-converting enzyme (ACE)

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 (C) UNCOMPETITIVE INHIBITION III) IRREVERSIBLE INHIBITION
Uncompetitive inhibition (UCI) only binds to the allosteric
Also known as suicide inhibitors
site of the enzyme-substrate complex
Similar to competitive inhibitor
o Allosteric site only opens up once the ES complex is
o Also a structural analog of the substrate
formed
o Hence, can also bind to the active site
Enzyme binds to the substrate → allosteric site opens up
Differs from competitive inhibitor in that it forms covalent
→ UCI binds to the allosteric site → ESI complex formation
bonds
→No products formed
o Covalent bonds – very strong bonds → difficult to be
When the inhibitor is released → ES complex formation
broken down → cannot be reversed by increasing the
→ products are formed (Figure 4)
substrate concentration
Inhibitor binds to the active site→ EI complex formation (via
(1) Effect on Vmax covalent bonds) → suicide inhibition (Figure 5)
↓ Vmax
o Not all enzymes can only be bound to the substrate
o Enzymes can form either ES complex formation or ESI
complex formation
o Can nerver reached Vmax
Some enzymes participate in the ESI complex formation →
↓ ES complex formed →↓ products formed Figure 5. Irreversible inhibition.

(2) Effect on Km (1) Examples

↓ Km (i) Sarin
Equilibrium is changed (shifted to the right)
Nerve gas
ESI formation → ↓ ES complex formation → reaction
Used in chemical warfare
shifts to the right (by Le Chatelier’s principle) → ↓
Inhibits acetylcholinesterase
substrate concentration → ↓ Km
o Acetylcholinesterase – breaks down acetylcholine

(ii) Aspirin
Irreversible cyclooxygenase (COX) inhibitor
 COX – catalyzes the formation of prostaglandins
and leukotrienes [Harvey & Ferrier, 2011]

Figure 4.Uncompetitive inhibition.

(3) Examples

(i) Arsenic
Poison
Inhibit glyceraldehyde-3-phosphate dehydrogenase
(G3PDH)
o G3PDH – converts glyceraldehyde-3-phosphate to
1,3-bisphosphoglycerate

(ii) Lithium
Inhibits an enzyme in the phosphoinositide pathway

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 IV) APPENDIX

Table 1. Summary of Reversible and Irreversible Inhibitors

Irreversible
Reversible Inhibitors
Parameter Inhibitors
Competitive Noncompetitive Uncompetitive
Binds with the Binds only with the Binds with the active
Binds with the active
allosteric site of either allosteric site of the site via covalent
(i) Mechanism site via noncovalent
the free enzyme or ES complex bonds
bonds
the ES complex
↓ -
(ii) Km ↑ Normal

↓ ↓ -
(iii) Vmax Normal

Statins MAO-I Arsenic Sarin

(iv) Examples ACE-I Oxamic acid Lithium Aspirin

Figure 6. Overview of enzyme inhibition.

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 V) REVIEW QUESTIONS

1) Which of the following statements is correct about

enzyme kinetics?
a) Km is a reflection of affinity
b) Km is the substrate concentration at half-maximal
velocity
c) Vmax is the velocity when all the enzymes are
saturated
d) All of the statements are correct.

2) Which of the following statements is correct about

competitive inhibition?
a) Reduces Km
b) Increases Vmax
c) Forms covalent bonds with the active site
d) Example are statins

3) Which of the following statements is correct about

noncompetitive inhibition?
a) Unchanged Km
b) Increases Vmax
c) Examples are ACE inhibitors
d) Binds only to the allosteric site of the ES complex

4) Which of the following statements is correct about

uncompetitive inhibition?
a) Increases Km
b) Decreases Vmax
c) Examples are MAO-I
d) Binds to the allosteric site of either the free enzyme
or the ES complex

5) Which of the following statements is correct about

irreversible inhibition?
a) Forms covalent bonds with the active site
b) Examples include aspirin and nerve gases
c) Increase in substrate concentration will not reverse
the inhibition
d) All of the choices are correct.

CHECK YOUR ANSWERS

VI) REFERENCES

Harvey, R., & Ferrier, D. (2011). Lippincott's Illustrated

Reviews - Biochemistry (5th ed.). Philadelphia: Lippincott Williams &
WIlkins.

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