Faculty of

Medicine
Medical
Education-
Damietta
University

Level 1
Semester 1
Module 1B
 Level 1
Semester 1
Module 1B
Introduction to Biochemistry
and
Basis of Genetics
Catalytic Proteins 4: Enzymes
Enzyme Regulation
• Contact: Medical Biochemistry Department.

• email: drosmanzaki@Hotmail.com

• Mobile: 01141512123

• Academic hours:
➢Sunday: 10:00-12:00 AM
➢Wednesday: 10:00-12:00 AM
 Contents

• Enzyme Regulation:

➢ Control of enzyme quantity:

• Altering the rate of enzyme synthesis and degradation.

• Induction.

• Repression.

➢ Altering the catalytic efficiency of the enzyme:

• Allosteric regulation.

• Feedback inhibition.

• Pro-enzyme (zymogen).

• Covalent modification.

• Protein-protein interaction.
 Learning outcomes
At the end of the lecture, the students should be
able to:

• Identify and describe different methods for

control of enzyme quantity.

• Identify and describe the different methods for

altering the enzyme catalytic efficiency.

• Correlate their knowledge to a clinical situation.

 Case scenario
(Clinical correlate)
• Amal, a 23-year old woman, 168 cm tall, is
being treated for anorexia nervosa. She
has been gaining weight, and is now back
to 62 kg from a low of 52 kg. Her blood
glucose is still below normal (fasting blood
glucose of 68 mg/dL, compared with a
normal range of 80-100 mg/dL). She
complains to her physician that she feels
tired when she jogs, and she is concerned
that the “extra weight” she has gained is
slowing her down.
 Regulation of enzyme activity
Why ?
❑ Regulation of enzyme activity is
important to coordinate the different
metabolic processes.

❑ It is also important for homeostasis i.e.

to maintain the internal environment of
the organism constant.
Achieved by two general mechanisms:
A) Control of enzyme quantity

➢ Altering the rate of enzyme synthesis and degradation.

➢ Induction.

➢ Repression.
B) Altering the enzyme catalytic efficiency:

➢ Allosteric regulation.

➢ Feedback inhibition.

➢ Proenzyme (zymogen).

➢ Covalent modification.
 Catalytic Protein
(Enzyme) Chemistry

ILO-1

Identify and describe different methods

for control of enzyme quantity.
A) Control of enzyme quantity
1- Control of the rates of enzyme synthesis
and degradation.
❖ Enzymes are protein in nature, they are
synthesized from amino acids under gene control
and degraded again to amino acids.
❖ Enzyme quantity depends on the rate of enzyme
synthesis and the rate of its degradation.
❖ Increased enzyme quantity may be due to an
increase in the rate of synthesis, a decrease in the
rate of degradation or both.
❖ Decreased enzyme quantity may be due to a
decrease in the rate of synthesis, an increase in the
rate of degradation or both.

* For example, the quantity of Liver Arginase

enzyme increases after protein rich meal due to
an increase in the rate of its synthesis.
 2- Induction
Induction means an increase in the rate of enzyme
synthesis by substances called inducers.
According to the response to inducers, enzymes are
classified into:
➢ Constitutive enzymes, the concentration of these
enzymes does not depend on inducers.
➢ Inducible enzymes, the concentration of these
enzymes depends on the presence of inducers.
▪ For example, induction of lactase enzyme in
bacteria grown on glucose media.
3- Repression

❑ Repression means a decrease in the rate

of enzyme synthesis by substances called
repressors.

❑ Repressors are low molecular weight

substances that decrease the rate of enzyme
synthesis at the level of gene expression.
❑ Repressors are usually end products of
biosynthetic reaction, so repression is
sometimes called feedback regulation.
❑ For example, dietary cholesterol decreases
the rate of synthesis of HMG CoA reductase
(β-hydroxy β-methyl glutaryl CoA reductase),
which is a key enzyme in cholesterol
biosynthesis.
4- De-repression

Following removal of the repressor

or its exhaustion, enzyme synthesis
retains its normal rate.
5- Concentration of substrates, coenzymes and
metal ion activator:
❑ The susceptibility of enzyme to degradation
depends on its conformation.
❑ Presence of substrate, coenzyme or metal ion
activator causes changes in the enzyme
conformation decreasing its rate of
degradation.
 Activity
• Enumerate ways to control quantity of
enzyme and discuss one of them

• Discuss feed back regulation (def,

mechanism ,example)
 Catalytic Protein
(Enzyme) Chemistry

ILO-2

Identify and describe the different

methods for altering the enzyme catalytic
efficiency.
B) Control of catalytic efficiency of enzymes

1- Allosteric Regulation

❑ Allosteric enzyme is formed of more than

one protein subunit.

❑ It has two sites; a catalytic site for substrate

binding and another site (allosteric site), that is
the regulatory site, to which an effector binds.
 Allosteric regulation

Allosteric means “other site”

Active site

E
Allosteric
site
❑ Allosteric means another site
❑ If binding of the effector to the enzyme increases it
activity, it is called positive effector or allosteric
activator e.g. ADP is allosteric activator for
phosphofructokinase enzyme.
❑ If binding of the effector to the enzyme causes a decrease in its
activity, it is called negative effector or allosteric inhibitor e.g.
➢ ATP and citrate are allosteric inhibitors for phosphofructokinase
enzyme.
➢ Glucose-6-phosphate is allosteric inhibitor for hexokinase
enzyme.
 The allosteric site the
enzyme “on-off” switch

Active
site

E Allosteric
Substrate site empty Conformational E
fits into change Inhibitor
the active molecule
Substrate
site is present
cannot fit
The inhibitor into the
molecule is active site Inhibitor fits
absent into allosteric
site
Mechanism of allosteric regulation
Binding of the allosteric effector to the regulatory site
causes conformational changes in the catalytic site, which
becomes more fit for substrate binding in positive effector
(allosteric activator), and becomes unfit for substrate binding in
negative effector (allosteric inhibitor)
 Allosteric regulation
• Conformational changes by regulatory molecules
– inhibitors
• keeps enzyme in inactive form
– activators
• keeps enzyme in active form

Conformational changes Allosteric regulation

2- Feedback Inhibition
➢ In biosynthetic pathways, an end product may
directly inhibit an enzyme early in the pathway.
➢ This enzyme catalyzes the early functionally
irreversible step specific to a particular biosynthetic
pathway.
➢ Feedback inhibition may occur by simple feedback
loop.
 Feedback Inhibition
– final product is inhibitor of earlier step

A→B→C→D→E→F→G
→
→

→
X
enzyme enzyme enzyme enzyme enzyme enzyme
1 2 3 4 5 6

End product is an inhibitor of enzyme 1

❑ Feedback inhibition can occur by multiple feedback inhibition
loops as occurs in branched biosynthetic pathways.

❑ Feedback regulation is different from feedback inhibition.

Feedback regulation:
➢ It means that an end product in the reaction
decreases the rate of enzyme synthesis at the
level of gene expression.
➢ It decreases the enzyme quantity through the
action on the gene that encodes the enzyme.
➢ It does not affect the enzyme activity.
➢ It is a complicated process that takes hours to
days.
➢ For example, inhibition of HMG CoA reductase
enzyme by dietary cholesterol.
Feedback inhibition
➢ It means that an end product directly
inhibits an enzyme early in biosynthetic
pathways.
➢ It does not affect enzyme quantity.
➢ It decreases the enzyme activity.
➢ It is a direct and rapid process that occurs
in seconds to minutes.
➢ For example, CTP inhibits aspartate
transcarbamylase enzyme in pyrimidine
synthesis.
 Activity
• 1- During ___________the final product of a metabolic pathway turn off the
first step of metabolic pathway.

(A) Positive feed back

(B) Negative feed back

(C) Competitive feed back

(D) Both A and C

• 2- An allosteric modulator influences enzyme activity by

(A) Competing for the catalytic site with the substrate

(B) Binding to a site on the enzyme molecule far from the catalytic site

(C) Changing the nature of the product formed

(D) Covalently modifying enzyme

 Activity
• Enumerate factors control catalytic activity
of enzyme and discuss one of them

• Discuss allosteric regulation (def,

mechanism ,types )

• Compare between feedback regulation and

feed back inhibition
3- Pro-enzymes (Zymogens)
➢ Some enzymes are secreted in inactive
forms called pro-enzymes or zymogens.
➢ Examples for zymogens include:
1. Pepsinogen,
2. Trysinogen,
3. Chymotrypsinogen,
4. Prothrombin and Clotting factors.
➢ Zymogen is inactive because it contains an additional
polypeptide chain that masks (blocks) the active site of the
enzyme.

➢ Activation of zymogen occurs by removal of the polypeptide

chain that masks the active site.
Activation of chymotrypsinogen to chymotrypsin,
and of trypsinogen to trypsin
 Activation of zymogens can occur:

a) Activation by HCl
HCl
Pepsinogen Pepsin

b) Auto activation i.e. the enzyme activates

itself.
Pepsin
Pepsinogen Pepsin
c) Activation by other enzymes

Enterokinase
Trypsinogen Trypsin

Thrombokinase + Ca++
Prothrombin Thrombin
Biological importance of zymogens

➢ Some enzymes are secreted in zymogen

from to protect the tissues of origin from
auto digestion.

➢ To insure rapid mobilization of enzyme

activity at the time of needs in response to
physiological demands.
4- Protein-protein interaction

❑ In enzymes that are formed from of many protein

subunits, the enzyme may be present in an inactive
from through interaction between its protein
subunits.
❑ The whole enzyme, formed of regulatory and
catalytic subunits, is inactive.
❑ Activation of the enzyme occurs by separation of
the catalytic subunits from the regulatory subunits.
• Protein kinase A enzyme is an example for
regulation of enzyme activity by protein interaction.
• It is formed of 4 subunits, 2 regulatory (2R) and 2
catalytic (2C) subunits.
• The whole enzyme (2R2C) is inactive.
• cAMP (cyclic adenosine monophosphate) activates
the enzyme by binding to the 2 regulatory (2R)
subunits releasing the 2 catalytic (2C) subunits.
5- Covalent modification

❑ It means modification of enzyme activity

through formation of covalent bonds e.g.

➢ Methylation (addition of methyl group).

➢ Hydroxylation (addition of hydroxyl group).

➢ Adenylation (addition of adenylic acid).

➢ Phosphorylation (addition of phosphate group).

 Reversible covalent modification
What’s covalently modulated
enzymes?
•Activity is modulated by covalent modification of one or
more of it’s amino acid residues in enzyme.
• Common modifying groups include: phosphoryl, adenylyl,
methyl and hydroxyl.

• These groups are generally linked to and removed from the

regulatory enzyme by separate enzymes.
➢ Phosphorylation is the most covalent
modification used to regulate enzyme activity.
➢ Phosphorylation of enzyme occurs by
addition of phosphate group to the enzyme at
the hydroxyl group of serine, threonine or
tyrosine.
➢ This occurs by protein kinase enzyme.
Protein kinases catalyze the
phosphorylation of proteins
➢ Dephosphorylation of the enzyme occurs by removal
of phosphate group from the hydroxyl group of
serine, threonine or tyrosine.

➢ This occurs by phosphatase enzyme.

 ➢The phosphorylated from is the active form in
some enzymes, while the dephosphorylated form
is the active form in other enzymes.

(active form) (inactive form)

(active form)
(inactive form)
Enzymes activated by phosphorylation:
❑ These are usually enzymes of degradative
(breakdown) reactions e.g.
1. Glycogen phosphorylase that breaks
down glycogen into glucose.
2. Lipase that hydrolyzes triglyceride into
glycerol and 3 fatty acids.
Enzymes inactivated by phosphorylation:

❑ These are enzymes of biosynthetic

reactions

1. Glycogen Synthetase, which catalyzes

biosynthesis of glycogen.

2. HMG CoA reductase, an enzyme in

cholesterol biosynthesis.
 Activity
1- The inactive precursor of an active enzyme is called

A) Zymogen

B) Ribozyme

C) Isozyme

D) Apoenzyme

2- Activation or inactivation of certain key regulatory enzymes is

accomplished by covalent modification of the amino acid:

(A) Alanine

(B) Lysine

(C) Phenylalanine

(D) Serine
 Activity
• Discuss protein- protein interaction ( mechanism
,one example )
• Discuss proenzyme and its biomedical
importance
• Enumerate methods of covalent modification of
enzyme activity
• Discuss covalent modification by
phosphorylation (types, enzymes involved,
examples)
 Catalytic Protein
(Enzyme) Chemistry
LO 3-

Correlation the knowledge to a clinical

situation
 Case Report and Clinical
Correlates
• One of the fuels used by Amal’s skeletal muscles
for jogging is glucose, which is converted to
glucose 6-P by the enzymes hexokinase and
glucokinase. Glucose 6-P is metabolized in the
pathway of glycolysis to generate ATP. This
pathway is feedback regulated, so that as her
muscles use ATP, the rate of glycolysis will
increase to generate more ATP.
Summary and rap up
Discussion
10 minutes
 Brain storming question

• What happened when Amal jogs? What does ATP use result in?

• When Amal jogs, the increased use of ATP for muscle contraction
results in an increase of AMP, which allosterically activates both the
allosteric enzyme phosphofructokinase1 and glycogen
phosphorylase. This is an example of feedback regulation by the
ATP/AMP ratio. Unfortunately, her low caloric consumption has not
allowed activation of the rate-limiting enzymes in her fuel storage
pathways, and she has very low glycogen stores. Consequently,
she has inadequate fuel stores to supply the increased energy
demands of exercise.
Feed back
 References

• Chatterjea’s Textbook of Medical Biochemistry,

8th edition.

• Vasudevan's Textbook of Biochemistry For

Medical Students, 6th Edition.

• BRS Biochemistry, Molecular Biology, and

Genetics, 5th Edition.