UTS:SCIENCE

Lecture 3: Part 3
Enzyme Regulation
Michael Widjaja

Metabolic
Biochemistry

Content

Reversible and irreversible inhibitors

Feedback Control
Allosteric Enzymes
Covalent Modification
Phosphorylation
glycogen phosphorylase

Activation by Cleavage
blood clotting

Metabolic
Biochemistry

Enzyme Inhibitors

Metabolic
Biochemistry

Irreversible inhibitors:
Dissociate very slowly from target enzyme due to tight
(covalent) binding
Drugs and toxins
Aspirin / Cyclooxygenase
Penicillin / Transpeptidase
Reversible Inhibitors:
Rapid dissociation of inhibitor from enzyme
Competitive
Uncompetitive
Mixed (non-competitive)
Used mainly in metabolism

Irreversible inhibitors

Metabolic
Biochemistry

Example: Penicillin
Antibiotic derived from yeast
Binds irreversibly to transpeptidase which results in an
unstable bacterialcell wall
Bacteria counter with -lactamase to inactivate penicillin
Now add clavulanic acid (-lactamase inhibitor)

Alexander Flemming
1929

Penicillium fungi

Reversible inhibitors

Metabolic
Biochemistry

Competitive inhibitors dock at enzyme active site

Substrate cannot bind
Product cannot be formed

Resembles substrate

Reversible inhibitors

Metabolic
Biochemistry

Uncompetitive inhibitors dock at another site on the

enzyme (allosteric site)
Only binds when enzyme and substrate are docked
(enzyme substrate complex)
Product cannot be formed

Reversible inhibitors

Metabolic
Biochemistry

Mixed (Non-competitive) inhibitors can bind free

enzyme or enzyme substrate complex
Product cannot be formed

Outcompeting inhibition

Metabolic
Biochemistry

Competitive inhibitors can be overcome by increasing substrate concentration

NOT- Outcompeting inhibition

Metabolic
Biochemistry

Mixed inhibitors cannot be overcome by increasing substrate concentration

Regulatory enzymes
Allosteric modulation
Regulation by cleavage
Regulation by covalent modifications

Metabolic
Biochemistry

Regulation
Living cell
Factory
Metabolic pathways
Assembly lines
Enzymes
Machine/ workers
Substrates
Raw materials
Catalysis
Processing
Products

Metabolic
Biochemistry

Feedback Control
2 types of regulation
Positive = more
Negative = less

1st enzyme in reaction

will be targeted for
regulation

Metabolic
Biochemistry

Metabolic
Biochemistry

Allosteric modulation
Allos = other; steric = shape or side

Where a modulator binds to a different part are of the

enzyme which is not the active site and leads to a change
in the shape or size of the active site affects binding of
substrate

Where allosteric enzymes often work

The first step is a good place for an inhibitor (red arrow)
Branch points in a pathway (green arrows)
F
A

E
H

Metabolic
Biochemistry

Allosteric modulation

E.g. Aspartate Transcarbamoylase (ATCase)

Converts aspartate (asp) and carbamoyl phosphate (CP) to
cytidine triphosphate (CTP)
Cytidine triphosphate binds to allosteric site of ATCase to
inhibit binding of aspartate and carbamoyl phosphate

CP

asp

Active site

ATCase

ATCase

Allosteric site
CTP

Metabolic
Biochemistry

Allosteric modulation
1. Positive Effector
allosteric site
effector

effector

structural
transition

Taut state
(less active)

Relaxed state
(more active)

effector

2. Negative Effector
effector
structural
transition

Allosteric modulation

Metabolic
Biochemistry

Allosteric enzymes are often multimeric

A group (complex) of subunits (single enzymes)

Binding to one subunit alters overall shape due to

domino effect
Homotropic allosteric enzymes
Effector as the substrate
Active site and regulatory site are same site

Heterotropic allosteric enzymes

Effector is not substrate for enzyme
Active site and regulatory sites are different

Allosteric enzymes- deviant behavior

Metabolic
Biochemistry

Michaelis-Menton plot
Regular enzyme:
hyperbolic-shape

v
Velocity

Allosteric enzyme:
Sigmoidal shape
[S]
Substrate Concentration

Autumn 2014

Copyright UTS 2014

17

Activation by cleavage

Metabolic
Biochemistry

Zymogens (Proenzymes)
Proenzymes are enzymes that need to be cleaved in order
to be active
Cleavage allows rapid activation

E.g. Trypsinogen (zymogen) cleaved into Trypsin

(enzyme)
Digests proteins (from meals) in small intestine
Needs to be activated

Activation by cleavage

Metabolic
Biochemistry

E.g. Clotting factors

Domino affect: one enzyme gets activated that activates
another

Activated
Enzyme

Zymogen

Metabolic
Biochemistry

An Enzyme Cascade
1 trigger molecule

5 enzymes
(zymogen
cleavage)
activated

Each of
these now
activates
5 other
zymogens

Regulation by covalent modification

Metabolic
Biochemistry

Includes:

Cleavage
Phosphorylation (addition of phosphate group)
Adenylation (addition of adenyl group)
Acetylation (addition of acetyl group)
Methylation (addition of methyl group)
Glycosylation (addition of sugar groups)

Reversible

Enzyme phosphorylation
Most common
Kinases phosphorylate
Add phosphate group

Phosphatases de-phosphorylate
Remove phosphate group

Autophosphorylation
Adds a phosphate group without any help
from kinases

Metabolic
Biochemistry