Allosteric Regulation
Allosteric Enzymes- some enzymes possess
additional sites (Allosteric sites) besides active
sites are called allosteric enzymes
The binding of allosteric modulators (small
signal molecules, often small metabolites
or cofactors) at specific sites (allosteric
sites) distinct from the active site triggers
conformational changes that are
transmitted to the active site
(intramolecular signal transduction)
Properties of Allosteric Enzyme
1. Catalyze essentially irreversible reactions; are rate
limiting
2. Generally contain more than one polypeptide chain
3. Do not follow Michaelis-Menten Kinetics
4. Are regulated by allosteric activators or inhibitors
5. Can be up-regulated by allosteric activators at constant
[S]
6. Can be down regulated by allosteric inhibitors at
constant [S]
7. Activators and Inhibitors need not have any structural
resemblance to substrate structure
8. Give a sigmoidal curve when the graph is plotted
between v and (S)
Allosteric enzymes are often oligomeric:
The aspartate transcarbamoylase (ATCase) consists
of two catalytic trimers and three regulatory dimers
CTP CTP
CTP CTP
T- state
Major conformational
differences were
observed for the
ATCase strucutures in
the presence of CTP
ATP ATP and ATP.
ATP ATP
The active site is ~50 A away
R-state from the allosteric site.
Classes
K- class: changes Km and not Vmax
e.g. phosphofructokinase
V- class: changes VMax and not Km
e.g. acetyl CoA carboxylase
Homotropic Effect- if substrate influences the binding
of the substrate through allosteric mechanism and it
is always Positive
Heterotropic effect- allosteric modulator affect
substrate binding and may be positive/ negative
Activation of latent Enzyme
Some enzymes (and other proteins) are
activated via proteolytic cleavage of one
or more peptide bonds of precursor
proteins (zymogens/ Proenzymes or
preproteins)
Many proteases activated this way can be
inactivated by inhibitor proteins tightly-
bound in the active sites
Active chymotrypsin
and trypsin are
produced from
inactive zymogens via
proteolytic cleavage,
with
Covalent Modification
• Regulation by covalent modification is slower
than allosteric regulation
• Reversible
• Require one enzyme for activation and one
enzyme for inactivation
• Covalent modification freezes enzyme T or R
conformation
Phosphorylation /dephosphorylation
•most common covalent
modification
• involve protein
kinases/phosphatase
•PDK inactivated by
phosphorylation
•Amino acids with –OH groups are
targets for phosphorylation
•Phosphates are bulky (-) charged
groups which effect conformation
Compartmentation
• Anabolic and catabolic pathways operates in
different compartment of cell
e.g. enzymes for fatty acid metabolism
Control of syntehsis
• Amount of enzyme directly control overall
rate of reaction
• Induction- increased expression of gene
• Repression- decreased expression of gene
• Mediated via hormones or other substances
Degradation
• Variability in half life
• Ranging from minutes to days
• Generally regulatory enzymes degrade
rapidly
Isozymes
• Multiple forms of the enzymecatalysing the
same reaction are isozymes. They differ in
their physical and chemical properties
which include the structure, electrophoretic
and immunological properties etc.
Abzymes-1
• Catalytic antibody, abzyme (mid-80s)
• Abzymes which are able to hydrolyze proteins, DNA, RNA, or
polysaccharides have been found in the sera of patients with
autoimmune and also viral pathologies
• Possible applications
– As sequence-specific cleaving agents to destroy peptides or
carbohydrates associated with viral particles or tumor cells
– The genetic deficiency of an extracellular enzyme could be cured
by immunization with an appropriate hapten to elicit catalytic
antibodies that would substitute the missing enzyme
– Catalytic antibodies with the capacity to degrade cocaine into the
resulting nontoxic products for cocain addiction
– Detoxification after accidental exposure to insecticides or the
intentional poisoning by nerve gas
Abzymes-2
• One of the most fascinating potential applications is in the area
of prodrug activation in cancer treatment using a strategy called
Antibody Directed Abzyme Prodrug Therapy (ADAPT)
