Enzyme Inhibition

Enzyme inhibition is defined as a substance

which binds with the enzyme and brings about a
decrease in catalytic activity.

The inhibitors may be drugs, antibiotics, toxins,

and antimetabolite.
 Types of inhibition
Reversible
Irreversible
1. Competitive inhibition – Reversible

2. Non – competitive – Reversible or

irreversible depending on type of
enzyme

3. Uncompetitive – Reversible

4. Suicidal inhibition (Irreversible)

 Competitive inhibitor
Inhibitor molecules
are competing with
the normal substrate
molecules for binding
to the active site of
the enzyme
Inhibitor is a
substrate analog
Inhibitor binds to
active site of enzyme.
Malonate inhibits succinate dehydrogenase enzyme
Competitive Inhibition

Typically, I is a substrate analog.

Km is increased in presence of
competitive inhibitor.
The affinity of the enzyme
towards substrate is apparently
decreased in presence of the
inhibitor.
Vmax is not changed
Competitive inhibition is usually
reversible.

Excess substrate abolishes the inhibition.

Clinical significance of
competitive inhibition
 Sulphonamides
Antibacterial agent
Bacterial wall is impermeable
PABA
to folic acid.
O
Bacteria synthesize folic acid
from PABA HO C NH2

Sulpha drugs, being structural

analog of PABA, will inhibit Sulfanilamide
the folic acid synthesis in
bacteria, and they die. O

The drug is nontoxic to human H2N S NH2

cells, because human beings O

cannot synthesize folic acid.

 Methotrexate
Anticancer agent
It is a structural
analogue of folic
acid,
Competitively
inhibit folate
reductase enzyme
This is essential for
DNA synthesis and
cell division.
 Dicoumarol
It is structurally similar to vitamin K

Act as an anticoagulant by competitively inhibiting the

vitamin K activity
Isonicotinic acid hydrazide (INH)
It is a commonly used antituberculous drug.

Structurally similar to pyridoxal

Prolonged use of INH may cause pyridoxal

deficiency and peripheral neuropathy
 Allopurinol

Used in the treatment of

gout
Structural analog of
hypoxanthine
 Competitive inhibitors
Drug Enzyme inhibited Clinical use
Allopurinol Xanthine oxidase Gout
Dicoumarol Vitamin K-epoxide reductase Anticoagulant
Penicillin Transpeptidase Bacteria
Sulphonamide Pteroid synthetase Bacteria
Trimethoprim Tetrahydro folate reductase Bacteria
Methotrexate Tetrahydro folate reductase Cancer
Pyrimethamine Tetrahydro folate reductase Malaria
6-Mercaptopurine Adenylosuccinate synthetase Cancer
5-fluorouracil Thymidylate synthetase Cancer
Azaserine Polyribosyl amidotransferase Cancer
Cytosine arabinoside DNA polymerase Cancer
Acyclovir DNA polymerase Virus
Neostigmine Acetylcholinesterase Myasthenia gravis
Alpha-methyl dopa Dopa-decarboxylase Myasthenia gravis
lovastatin HMG-CoA reductase Cholesterol
 Non competitive – Irreversible inhibition
The inhibitor usually
binds to a different
domain on the
enzyme, other than the
substrate binding site.
There is no
competition between
substrate and inhibitor.
There is no structural
resemblance with the
substrate
The inhibitor combines with the enzymes by forming a
covalent bond and then the reaction becomes
irreversible.
Increasing the substrate concentration will not abolish
non-competitive inhibition.
Vmax is reduced
Km is not changed
 Clinical significance
1. Cyanide : Inhibits cytochrome oxidase.

2. Fluoride : Inhibits the enzyme ,enolase, and

consequently glycolysis.

3. Iodoacetate : Inhibit enzymes having-SH

group in their active centers.

4. BAL (British Anti Lewisite; dimercaprol) : is

used as an antidote for heavy metal poisoning.
Difference between competitive & noncompetitive
inhibition
Competitive Noncompetitive
inhibition inhibition
Acting on Active site Other than active site
Structure of Substrate analogue Unrelated molecule
inhibitor
Inhibition Reversible Generally irreversible
Excess substrate Inhibition relieved No effect
Km Increased No change
Vmax No change Decreased
significance Drug action Toxicological
 Uncompetitive inhibition
The inhibitor does not have any
affinity for free enzyme.
Inhibitor binds to enzyme–substrate
complex; but not to the free
enzyme.
Both Vmax and Km are decreased
Inhibition of ALP by
phenylalanine. (Regan isoenzyme)
by phenylalanine is an example of
uncompetitive inhibition.
 Suicide inhibition
Irreversible inhibition of enzyme activity.

The inhibitor makes use of the enzyme's own

reaction mechanism to inactivate it

Inhibitor is a structural analog

Initially it binds to the enzyme that is to be
inhibited and gets converted to a more effective
product

The new product irreversibly binds to the

enzyme and inhibits further reactions.
 Allopurinol Aspirin
A strong inhibitor of Anti-inflammatory drug.
xanthine oxidase gets Prostaglandins are responsible for
oxidized to alloxanthine. inflammatory reactions by the
enzyme Cyclo-oxygenase
In the treatment of GOUT
Aspirin binds to the active center of
. cyclo-oxygenase, and prostaglandin
synthesis is inhibited, and
inflammation subsides
Enzyme regulation
In a metabolic pathway catalyzed by series of
enzymes activity , one enzyme regulates the
over all rate of metabolic sequences as per the
cells demand

Such enzymes are called regulatory enzymes

 Types of enzyme regulation
1. Allosteric regulation
2. Feed back regulation
3. Covalent modification
4. Induction and repression
5. Regulation by proteolytic cleavage
 Allosteric regulation
Enzymes possess additional sites, known as
allosteric sites
Allosteric enzyme has one catalytic site where
the substrate binds and another separate
allosteric site where the modifier binds
Positive modifier: enhance
the activity of the enzyme
(allosteric activation)

Negative modifier: inhibit

the activity of the enzyme
(allosteric inhibition).
Co-operativity :The
binding of substrate
to one of the subunits
of the enzyme may
enhance substrate
binding by other
subunits.
Pathway Enzyme Activator Inhibitor
Glycolysis PFK 1 AMP ATP & Citrate

TCA cycle Isociterate ADP ATP

dehydrogenase
Glycogenolysis Glycogen Phosphorylase AMP ATP

Gluconeogenesis Fruc 1,6 Bisphosphatase ATP & Citrate AMP

Fatty acid Acetyl CoA Carboxylase Citrate

synthesis
 Feed back inhibition
The activity of the
enzyme is inhibited
by the final product
of the biosynthetic
pathway.
 Covalent modification
Either addition of a group to the enzyme
protein by a covalent bond; or removal of a
group by cleaving a covalent bond.
Most frequently addition of phosphate group –
phosphorylation
Removal of phosphate group –
dephosphorylation
Glycogen phosphorylase is a muscle enzyme
that breaks down glycogen to provide energy.
This enzyme exists in two interconvertible
forms.
Phosphorylase b (dephospho enzyme) is
inactive which is converted b y
phosphorylation of serine residues to active
form phosphorylase a.
 Proteolytic cleavage
Some enzymes are synthesized as Proenzymes or zymogens
which undergo irreversible covalent activation by the
breakdown of one or more peptide bonds.
Proenzymes namely chymotrypsinogen pepsinogen and
plasminogen, are respectively- converted to the active
enzymes chymotrypsin, pepsin and plasmin
 Induction & repression
Induction is effected through the process of
derepression.
The inducer will relieve the repression on the
operator site and will remove the block on the
biosynthesis of the enzyme molecules.
Tryptophan pyrolase and transaminases are
induced by glucocorticoid.
Glucokinase is induced by insulin.
ALA synthase is induced by barbiturates.
 Repressor
Both inhibition and repression reduce the
enzyme velocity, but the mechanisms are
different.
In the case of inhibition, the inhibitor acts on
the enzyme directly and the inhibitory activity
is noticed and the number of enzyme molecules
is not changed by the inhibitor.
Repressor acts at the gene level; and the
number of enzyme molecules is reduced in the
presence of repressor molecule.
The key enzyme of heme synthesis, ALA synthase is
autoregulated by the heme by means of repression.
The structural gene is transcribed and later translated to produce
the enzyme molecules. The transcription process starts at the
operator site when it is free. When heme is not available, this
operator site is open, and therefore the enzyme is being
synthesized.
When heme is produced in plenty, heme acts as the co-repressor
and in combination with an apo-repressor, heme will shut off the
operator site. Now further production of ALA synthase is stopped.
 Compartmentalization
Certain enzymes of the pathway may be
located in mitochondria whereas certain other
enzymes of the same pathway are cytoplasmic.
Example :
Heme synthesis
Urea cycle
Gluconeogenesis