chap t er

Enzymes Regulation

17

Learning ObjectivesExam Index

Understanding why regulation is needed

Basic classification of regulatory mechanisms
Effect of various enzyme regulations on enzyme kinetics
Understanding the role of inhibitors developing drugs and studying biology

17.1. Introduction

Interesting Fact
As we learnt in previous chapters that enzymes are pivotal to biochemical
reactions, their roles in driving the metabolism is indispensable. However, a cell has
Arslan et al. introduced intramolecular crosslinks
enormously complicated network of pathways and therefore it is in order to keep
between two domains of the Escherichia coli
the metabolic flux balanced, the enzymes which are catalysing those reaction must
helicase Rep, which unwinds DNA. By inserting
be regulated. (Otherwise condition would be like driverless cars running without
linkers of different lengths, the domains can
control causing chaos). This chapter on enzymology provides a comprehensive
be held either open or closed. The closed
information on regulation of enzymes. Understanding regulation of enzyme has
conformation activates the helicase, but it
become clinically important domain of science. As most of the enzymes are prime
can also generate super-helicases capable of
targets for developing drugs which either inhibit or augment the enzymatic activities.
unzipping long stretches of DNA at high speed
and with considerable force. Comstock et al. used
It is therefore very important to focus on fundamentals of enzyme regulation and to
optical tweezers and fluorescence microscopy to
understand alterations in their kinetics during such regulation. The cell has several
simultaneously measure the structure and function
mechanisms to regulate enzyme activity, first and the generalised mode (global
of the bacterial helicase UvrD. They monitored its
regulation) of regulation is compartmentalization of metabolism (discussed in
DNA winding and unwinding activity and its shape
chapter 13), usually the anabolic and catabolic compartments are separate and
during these activities. The motor domain also has
enzymes have isoforms with different kinetics abilities in various compartments.
a closed conformation during DNA unwinding and
The second mode of regulation of enzyme activity is availability of accessory
switches to a reversed open conformation during
the zipping-up interaction (Sciecne, 2015 (348)
factors (co-factors) for the enzymes, which are either obtained exogenously
(vitamins) or produced endogenously. The Third mode of regulation is via regulation
of substrate concentration, if there is no substrate, no product will be formed.
However, the above modes do show enzyme specificity. There are some specific routes of enzyme regulation which specifically modulate
enzyme thereby affecting its activity. In the following section we will consider such specific regulatory mechanisms.
Trick to Remember
Regulation of enzymes is like controlling electrical devices in household or industry. For safety purpose there are some general
cut-outs (like MCBs) and centralised switches which can switch off the whole power connection of a building (analogous to
global regulation), while for specific regulation of an electrical appliance we have individual switches (analogous to specific
regulation). Sometimes those devices which are frequently used and need to be operated from multiple places may have
multiple switches like a bulb on staircase may have more than one switch (on each floor) or room light may have additional
switch close to bed, similar to those enzymes which are regulated (analogous to allosteric regulation) .

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Chapter 17: Enzymes Regulation 335

17.2. Types of Enzyme regulation

Although there are several ways to classify the regulatory mechanisms of enzymes but based on their kinetics studies and the
basic distinction in mechanism of regulation we may organize enzyme regulation into four major classes. 1. Allosteric regulation,
2. Regulation by covalent modification, 3. Regulation by peptidal cleavage, 4. Regulation by selective inhibitor.

Modes of Enzyme Regulation

Allosteric
Regulation

Homotrophic

Regulation by
Covalent Modification

Reversible

Heterotrophic

K-Type

Regulation by
Peptidyl Cleavage

Regulation by
Peptidyl Cleavage

V-Type

Competitive

Irreversible

Mixed

Uncompetitive
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Fig. 17.1. Classification of major types of enzyme regulation processes (*Noncompetitive is a special case of Mixed)

17.3. Allosteric regulation

In biochemistry, allosteric regulation is the regulation of an enzyme or other protein by binding an effector molecule at the proteins
allosteric site (that is, a site other than the proteins active site). Effectors that enhance the proteins activity are referred to as
allosteric activators, whereas those that decrease the proteins activity are called allosteric inhibitors. The term allostery comes
from the Greek word allos which means other and stereos which means solid object. Allosteric regulation is the regulation of
an enzyme or other protein by binding an effector molecule at the proteins allosteric site (i.e., a site other than the proteins active
site). Effectors that enhance the proteins activity are referred to as allosteric activators, whereas those that decrease the
proteins activity are called allosteric inhibitors. The allosteric enzymes are usually capable of switching between active and
inactive depending on the state of the enzyme, regulators can be used to stabilize the enzymes conformation. The basic principle
of allosteric regulation is the binding of an activator at regulator site which can stabilize the active form of that allosteric enzyme,
while the binding of an inhibitor which stabilizes the inactive form of the enzyme. In the cell, activators and inhibitors dissociate
when at low concentrations, which then allows the enzyme to transit between active and inactive forms. This alteration in the
activity or concentration of regulators can cause a sophisticated pattern of response in the activity of cellular enzymes.
Before we discuss more on allosteric enzyme let us first understand meaning of active site and allosteric site.

17.3.1. Active site and allosteric site

Active site of an enzyme is defined as the region of an enzyme where substrate molecules bind and undergo a chemical reaction.
An active site may consist of two basic components formed by weak interaction between the side chain of various residues, one
called as binding site which facilitate the direct interaction between substrate and enzyme by offering the three dimensional
surface as well interacting groups (polar or non-polar depending on the kind of interaction). The second component of the active
site is catalytic site where reactant is converted into product. Most of the time binding site and catalytic site share steric space
in enzyme.

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336 Concise Chemistry

In general appearance, the active site is usually a groove or pocket of the enzyme which can be located in a deep tunnel within
the enzyme or between the interfaces of multimeric enzymes. There are two proposed models of how enzymes fit to their specific
substrate: the lock and key model and the induced fit model (Fig 17.2).
Key and lock hypothesis: Emil Fischers proposed the lock and key model, this assumes that the active site is a perfect fit
for a specific substrate and that once the substrate binds to the enzyme no further modification occurs. However, this hypothesis
could not explain the extra-stability of transition state.
Induced fit hypothesis: This was a modified version of key and lock hypothesis given by Daniel Koshland. The induced fit
model is a development of the lock-and-key model and assumes that an active site is flexible and it changes shape until the
substrate is completely bound. The substrate is thought to induce a change in the shape of the active site. The hypothesis also
predicts that the presence of certain residues (amino acids) in the active site will encourage the enzyme to locate the correct
substrate. Conformational changes may then occur as the substrate is bound.
Lock and Key Model (Emil Fischer)
Enzyme

Induced Fit Model (Koshland)

Enzyme

ES complex

Substrate

ES complex

Substrate

Transition state
structure
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Fig. 17.2. Difference between Key and Lock hypothesis and induced fit hypothesis

17.3.1. Key features of Allosteric enzymes

Historically, the primary studies on allosteric enzymes were performed on aspartate transcarbamylase (ATCase). Aspartate
carbamoyltransferase (or aspartate transcarbamylase, ATCase) plays a central role in the regulation of the pyrimidine pathway
in bacteria. The Holoenzyme is a dodecamer composed of six catalytic chains, each with an active site, and six regulatory chains
lacking catalytic activity. The catalytic subunits exist as a dimer of catalytic trimers, (2xC3), while the regulatory subunits exist as
a trimer of regulatory dimers, (3xR2), therefore the complete holoenzyme can be represented as (C3)2(R2) 3. The association of the
catalytic subunits C3 with the regulatory subunits R2 is responsible for the establishment of positive co-cooperativity between
catalytic sites for the binding of aspartate and it dictates the pattern of allosteric response toward nucleotide effectors. Fig 17.3
illustrates basic strategy to demonstrate the allosteric nature of the enzyme.
NATIVE SDS+BME

Native protein

Add substrate and

accessory factors
Isolated
and Purified

catalyzed the reaction

(Sigmoidal Kinetics)
Affected by adding CTP and ATP

catalyzed the reaction

Kinetics
(Micahelis menten type)
NO effect of adding CTP or ATP

Isolated
and Purified
Electrophoresis

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NOT catalysed the

reaction
but binds to modulators

C3

C3

Catalytic Unit
(Dimer of Trimers)
R2

R2

R2
Regulatory Unit
(Trimer of Dimers)

Fig. 17.3. Basic experiment to illustrate the salient features of allosteric enzymes using ATCase as model

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Chapter 17: Enzymes Regulation 341

General Reaction- Enzyme 1 ATP -----------------------> ENZ-PO4 1ADP

Key residues involved: Ser, Thr, Tyr, His
Example: In Glycogen Phosphorylase (undergoes phosphorylation at Ser14), phosphorylation converts less active
form b to more active form a. While in case of Glycogen synthase the phosphorylation causes conversion of an active
enzyme (a form) into inactive (b form) (Refer chapter 14 for details).

17.4.2. Regulation by Adenylation

Adenylylation is also now known as AMPylation, as it involves the addition of adenosine monophosphate (AMP) molecule
covalently to a protein side chain, altering the function of the protein. Adenylylation involves a phosphodiester bond between a
hydroxyl group of the molecule undergoing adenylylation and the phosphate group of the adenosine monophosphate nucleotide
(i.e. adenylic acid). This process can occur to molecules such as tyrosine residues. Enzymes that are capable of catalyzing this
process are called AMPylators.
Key residues: Tyr, Ala
General Reaction: Enzyme 1 ATP -----------------------> ENZ-AMP 1 PPi
Example: Glutamine synthase is activated by adenylation at Tyrosine residue.

17.4.3. Regulation by Uridylation

Similar to adenylation, Uridine nucleotides interact with proteins and causes the linkage via covalent medication at phosphodiester
linkage. Uridylation, involves a phosphodiester bond between a hydroxyl group of the molecule undergoing uridylation and the
phosphate group of the Uridine monophosphate nucleotide. Uridylation is controlled by cellular levels of alpha--ketoglutarate and
ATP. Apart from regulation of enzyme activity and modification of proteins, Uridylation is also commonly obsevered in RNA and
it mediated the degradation of RNA
General Reaction - Enzyme 1 UTP -----------------------> ENZ-UMP 1 PPi

Clinical Note

Bacterial ADP-ribosylating exotoxins

(bAREs) covalently transfer an ADP-Ribose
1 to target proteins of infected
moiety
of NAD
Example: PII protein that caused the adenylation of Glutamine synthase
is itself
regulated
by Uridylation. Additionally,
eukaryotes,
to yield
nicotinamide
and are
a
The Nitrogen regulatory protein P-II family contains a series of homologous
prokaryotic
signalling
proteins which
free
hydrogen
ion.
Upon
activation,
bAREs,
involved in the regulation of nitrogen metabolism. The proteins are post translationally modified via addition of a uridylyl
ADP-ribosylate any number of eukaryotic
group by the enzyme Uridylyl transferase.
proteins, such as GTP-binding proteins.
Subsequent activation of intracellular cyclic
AMP stimulates the release of fluid and ions
17.4.4. Regulation by ADP Ribosylation
from intestinal epithelial cells (resulting in
ADP-ribosylation is the addition of one or more ADP-ribose moieties to
vomiting and diarrhoea). There are a variety of
a protein. The source of ADP-ribose for most enzymes that perform this
bacteria which employ bAREs based infection
modification is the redox cofactor NAD1. In this transfer reaction, the
mechanism, eg. cholera of Vibrio cholera;
N-glycosidic bond of NAD1 that bridges the ADP-ribose molecule and the
heat-labile enterotoxin of E.coli; Exotoxin A
nicotinamide group is cleaved, followed by nucleophilic attack by the target
of Pseudomonas aeruginosa; Pertussis toxin
amino acid side chain. ADP-ribosyltransferases can perform two types of
of B. pertussis; C3 toxin of C. botulinum; and
modifications: mono-ADP ribosylation and poly-ADP ribosylation.
Diphtheria toxin of Corynebacterium diphtheria.

Key residues: Tyr

This is one of the common mode of enzyme regulation in prokaryotes

General Reaction - Enzyme 1 NAD -----------------------> ENZ-ADP Ribose 1Nicotinamide
Key residues: Glu, Arg, Cys, His
Example: Reversible ADP ribosylation of dinitrogenase reductase and Glutamine synthase, Regulation of DNA
ligase activity by poly (ADP-ribose)

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342 Concise Chemistry

Trick to Remember
Similar to the allosteric modulation, covalent modifications are also widespread in proteins which are not enzymes. As
covalent modification is also the common mode to activate or deactivate a non-enzyme protein such as transporter,
receptor or immune effector. Therefore examples here may include some non-enzyme proteins.

Tables 17.2 represents some of the commonly known protein-modifications their donor, biological roles and also the residues that
are prone to specific modification (some of them are examples of proteins but not enzymes)
Table 17.2. Commonly known covalent modifications and their examples.
Modification

Donor molecule

Example of modified protein

Protein function

Phosphorylation

ATP

Glycogen phosphorylase

Glucose homeostasis

Acetylation

Acetyl CoA

Histones

DNA packing

Myristoylation

Myristoyl CoA

Src

Signal transduction

ADP-ribosylation

NAD

RNA polymerase

Transcription

Farnesylation

Farnesyl pyrophosphate

Ras

Signal transduction

Thrombin

Blood clotting

g -Carboxylation

HCO3

Sulfation

3PAP*

Fibrinogen

Blood-clot formation

Ubiquitination

Ubiquitin

Cyclin

Control of cell cycle

*39-Phosphoadenosine59-phosphosulfate

Table 17.3 summarises additional modification on proteins (Some of them are non-enzymatic proteins and key residues on which
these modifications occur)
Table 17.3. Key covalent modifications on proteins and amino acids involved.
Name
Phosphorylation

Group added
-PO3

2-

Amino Acid Modified

S,T,Y

Examples
Regulatory enzymes,
Receptors

Acetylation

-CH2COO-

Histones

Methylation

-CH3

K,R

Histones

Acylation

Palmitic acid

G-protein-coupled receptors

Prenylation

Prenyl group

Ras p21

ADP-ribosylation

ADP-ribose

H,R

AMP

Adenylylation

G protein, eEF
Glutamine Synthetase

17.5 Enzyme regulation by limited proteolysis cleavage

While most of the enzymes and proteins in the cells are synthesized in their active form (e.g., the house-keeping enzymes and
structural proteins), a small but significant number of enzymes and proteins are produced as inactive precursors, which play
critical roles in cellular development and cell survival. These inactive precursors are known as zymogens or pro-proteins and

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344 Concise Chemistry

Fig. 17.6. Mechanism of zymogen activation of Trypsin and Chymotrypsin

17.6 Enzyme regulation by selective inhibitors

The inhibitors of enzyme activity are chemical substances, which in small quantity decrease the activity of enzymes in a specific
chemical way. As a result of the inhibitor - enzyme interaction enzyme-inhibitor complex is formed: once bound, the enzyme
cannot convert the EI complex to products. The existence of specific naturally occurring enzyme inhibitors, like antithrombin,
ant pepsin and antitrypsin, controls the enzyme activity in the human body and under physiological circumstances assures their
intracellular and extracellular action. Among the naturally occurring enzyme inhibitors there are also intermediary products
formed during some metabolic pathways. Product inhibition provides a limited mean of control or modulation of substrate flux
through the pathway. If one or more enzymes are allosteric enzymes particularly sensitive to product inhibition, the output of end
product of the pathway will be suppressed.
Enzyme regulation by inhibitors may be broadly grouped as reversible and irreversible inhibition, one which could be revered by
addition of more substrate is known as reversible inhibition while other which cannot be reversed until new molecules of enzymes
are synthesized are called irreversible inhibition.
Below is the description of some of the commonest modes of enzyme regulation by inhibitors.

17.6.1. Reversible Inhibition

In case of reversible Inhibition, inhibitors do not form any covalent bond with the enzyme and reaction can reverse back if
substrate concentration exceeds far from the inhibitor which means even if the reaction is completely inhibited by adding a
large amount of inhibitor, we can add large amount of substrate and product formation will begin. Reversible nature of these
reactions is also the basis of developing antidotes for various enzyme inhibitors. Based on the mechanism of the reaction there
are three basic type of reversible inhibitions, a. competitive inhibition, uncompetitive inhibition and mixed inhibition. Although
some biochemistry books mention non-competitive inhibition as fourth type, but we will realise in the flowing discussion that it
creates confusion (especially between uncompetitive and non-competitive).

Online Support
Video Lecture Available: You tube channel: video 17.2: Various types of reversible enzyme inhibition.

A. Competitive Inhibition
Competitive inhibition, as the term suggests, indicate the competition between substrate and inhibitor for the same binding site
on the enzyme. Substrate on binding to active site forms product, but inhibitor on binding with same site leads to formation of
non-productive EI complex. Increasing concentration of EI complex would inhibit the product formation more severely due to
a decrease in ES complex. Now the kinetics of EI formation is same as that of ES formation hence, the rate constant for the
formation of EI complex is given by Ki and a term alpha (a ) is introduced which represents the strength of inhibition (a 5 11
[I]/K I), where [I] is inhibitor concentration and Ki is the equilibrium constant for the breakdown of EI complex back to E and I,
hence with increasing inhibitor concentration the value of alpha would increase (Fig 17.7)

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Chapter 17: Enzymes Regulation 345

Salient features of competitive inhibition are:

Substrate competes with the inhibitor for the same site on the enzyme
Both substrate and inhibitor bind to the same site
EI complex do not lead to catalysis
Apperant Vmax same as V max (Not affected)

V + [S ]
) (alpha is define in text
Apperant Km Increased by a factor a or Km (apperant) 5 a Km (Equation: Vo = max
a
+
K
S
[
]
m
above)

Examples: Inhibition of Succinate dehydrogenase by malonic acid

Special case of Ethanol antidote in methanol poisoning

Inhibitor
Competition
for same site

=2

=1
No Inhbitor

ES

EI
NON-PRODUCTIVE

Vmax unchanged
Km increases

1
[S]

Increasing inhibitor
conc.

EI

=3

Vmax constant

Rate of Reactions

Ki

E+P
1
Vo

E+S
ES
+
1+ [I]
I = Ki

Hyperbolic plot
Increasing inhibitor
conc.

Double Reciprocal plot

Mechanism of inhibition

Km increases
Substrate concentration

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Fig. 17.7. Illustration of competitive inhibition a. Mechanism b. Lineweaver Burk plot c. Michaelis plot

B. Uncompetitive Inhibition
Uncompetitive inhibition, on the other hand, does not involve the competition with substrate, but the inhibitor binds to enzyme
substrate complex leading to an unproductive ESI complex. Infact, this occurs due to a conformational change in the enzyme
after binding of substrate which created a new binding site for inhibitor. So in this type of inhibition one would not observe the
EI complex in the reaction mixture. Similar to competitive inhibition the strength of inhibition in this case is indicated by similar
parameter (a 5 11 [I]/KI), where [I] is inhibitor concentration and Ki is the equilibrium constant for the breakdown of EI
complex back to E and I, hence with increasing inhibitor concentration the value of a would also increase (Fig 17.7).
Salient features of competitive inhibition are:
Inhibitor Binds to a site different than that of the binding site of substrate
It actually binds to ES complex to form non-functional ESI complex
ESI complex don not lead to catalysis
Apperant Vmax Vmax/a [Vmax is reduced] (a 5 11 [I]/KI) where [I] is inhibitor concentration and Ki is the
equilibrium constant for the breakdown of ESI complex
[Km is also reduced]
Apperant Km - Km / a 
V + [S ]
Equation: Vo = max
'S
Km +a[ ]
Example: The membrane-bound Na1/K1 ATPase is inhibited by ouabain, a toxic glycoside.

C. Mixed Inhibition
Competitive inhibition, is the mechanistic combination of competitive and uncompetitive inhibition, in this mode the inhibitor binds
to enzyme and forms EI complex, as well as it also binds to additional site created at non-catalytic site after binding of substrate
to enzyme, thus forming ESI complex. Therefore one can observe both EI and ESI complex in a reaction mixture operating by this
mechanism. The kinetics of this inhibition will also be a combination of above two, Infact the degree of EI formation or ESI formation
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346 Concise Chemistry

EI

=3
=2
=1

S
E

ES

No Inhbitor

Site for inhibitor

Vmax decreases

Inhibitor

ESI
NON-PRODUCTIVE

Km decreases

1
[S]

Vmax decreases

Increasing inhibitor
conc.

1+ [I]
Ki

E+P

Rate of Reactions

Ki

ES
+
I

1
Vo

E+S

Hyperbolic plot
Increasing inhibitor
conc.

Double Reciprocal plot

Mechanism of inhibition

Km decreases
Substrate concentration

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Fig. 17.8. Illustration of uncompetitive inhibition a. Mechanism b. Lineweaver Burk plot c. Michaelis plot

will be decided on the basis of values of a or . This gives rise to three

different conditions discussed in the following section.
Salient features of competitive inhibition are:
Inhibitor Binds to a site different than that of the binding site of
substrate or same site as well.
It actually binds to ES complex to form non-functional ESI complex
as well as EI complex
EI, ESI complex do not lead to catalysis
Apperant Vmax Vmax/a [Vmax is reduced]; ( 5 1 1 [i]/KI) and
( 5 1 1 [i]/KI)

Apperant Km a Km / a
[Km is increased when a > a ; Km is decreased when a <a ; and
remains unchanged when a 5a ]
V + [S ]
Equation: Vo = max
'S
aK m + a [ ]
Examples: L Threonine Dehydratase is inhibited by end
product isoleucine

Trick to Remember
As a smart trick to remember whether Km
or Vmax change in which type of inhibition,
an analogy may be created. In case of
competitive inhibition the lines on double
reciprocal plot are intersecting while on
a plot of uncompetitive lines are parallel
(Quite obvious! things that compete must
intersect and those which do not compete
remain parallel). So graph may be plotted
from this point and every new line placed
away from origin indicate increasing inhibitor
conc. Now recall the determination of Vmax
and Km on double reciprocal plot (chapter
16) and determine if Km and Vmax are
increasing or decreasing)

Mixed inhibition can give rise to three conditions

We have learnt above that mixed inhibition is mechanistically combination of both competitive and uncompetitive therefore the
kinetics and hence pattern of the graph will largely depend upon rate constants of EI formation and ESI formation (KI and KI)
and therefore the values of a and a . In the above figure the value of a < a so predominantly EI is formed. While if a > a
predominantly ESI complex will be formed and the graph will appear as below. As a special case when a 5 a the lines will
appear to merge at X axis and then Km will remain same. , such inhibition is called as NON competitive. This is a special
case of mixed and different from uncompetitive inhibition. No ESI complex would be formed in uncompetitive, but
here in non-competitive both EI and ESI complexes will attain similar equilibrium.

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Chapter 17: Enzymes Regulation 347

Double Reciprocal plot

EI

S
E

ES

No Inhbitor

Site for inhibitor

1+ [I]
Ki

Inhibitor

1+ [I]
Ki

Vmax decreases

EI

ESI

Km increases or decreases
or remains unchanged (depends on and )

NON-PRODUCTIVE

Increasing inhibitor
conc.

EI

Ki

E+P

Vmax decreases
Rate of Reactions

Ki

ES
+
I

1
Vo

E+S
+
I

Hyperbolic plot
Increasing inhibitor
conc.

Mechanism of inhibition

Km decreases

1
[S]

Substrate concentration

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No Inhbitor
Vmax decreases
Km decreases with increasing inhibitor

1
[S]

No Inhbitor

Also Known as
NON-COMPETITIVE
INHIBITION

Vmax decreases
Km decreases with increasing inhibitor

1
[S]

Increasing inhibitor
conc.

Mixed Inhibition (Condition 3)

= EI and ESi formed at same pace
1
Vo

Increasing inhibitor
conc.

Mixed Inhibition (Condition 2)

< ESI Complex predominates
1
Vo

1
Vo

Mixed Inhibition (Condition 1)

> EI Complex predominates

Increasing inhibitor
conc.

Fig. 17.9. Illustration of mixed inhibition a. Mechanism b. Lineweaver Burk plot c. Michaelis plot

No Inhbitor
Vmax decreases

Km Unchanged

1
[S]

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Fig. 17.10. Illustration of three conditions during mixed inhibition, the last curve shows non- competitive inhibition

17.6.2. Dixon plots: Inhibitor constant and rate of reaction

A graphical method for determination of the type of enzyme inhibition and the dissociation constant (Ki) for an enzyme-inhibitor
complex. The effect on the enzyme rate (V) is determined at two or more substrate concentrations, and over a range of inhibitor
concentrations (I). In a plot of 1/V against I, data for each substrate concentration fall on straight lines that intersect at I5-Ki and
1/V51/Vmax (competitive inhibition), or that intersect on the abscissa (1/v50) at I5-Ki (non-competitive inhibition).
The inhibitor constant, Ki, is an indication of how potent an inhibitor is; it is the concentration required to produce half maximum
inhibition.
Plotting 1/v against concentration of inhibitor at each concentration of substrate (the Dixon plot) gives a family of intersecting
lines.

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Concept Map

350 Concise Chemistry

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Chapter 17: Enzymes Regulation 351

Q1. A Protein moved more slowly is an SDS- PAGE. Isoelectric focusing [IEF] showed that here was no change in the
pI Mass spectrometric analysis showed that the modification was on serine The modification that the protein
undergoes is likely to be
[CSIR-NET DEC 2015]
a. Phosphorylation

b. Glycosylation

c. Ubiquitination

d. ADP- ribosylation

Solution: This question may be related to the activity of enzymes [by possible mode of protein modification],
however solving this does not require enzymology, but basic knowledge of proteins, Phosphorylation, Ubiquitination
and ADP ribosylation will cause a change in the ionization state of the protein and also the isoelectric point so the
separation on the pI will alter if there is any such mutation. But if modification is glycosylation the protein will
show a change in SDS PAGE but not in IEF. Therefore option b is correct.
Q2. Reaction products inhibit catalysis in enzymes by [CSIR NET JUNE 2013]
a. Covalently binding to the enzyme.

b. Altering the enzyme structure.

c. Occupying the active site.

d. from a complex with the substrate.

Solution: Reaction product often interact with the enzyme in many ways as stated in option a, c, and d, so correct
option may be b. Altering the enzyme structure.
Q3. Allosteric enzymes are

[GATE]

a. larger than simple enzyme

b. smaller than simple enzyme

c. larger and more complex than simple enzyme

d. smaller than simple enzyme but not complex

Solution: Allosteric enzymes are generally larger and more complex than simple enzymes (refer text for details)
Q4. Which one of the following statements is true about non-competitive inhibition?
a. Km increases

b. Km decreases

c. Vmax increases

d. Vmax decreases

[IISc, 2012]

Solution: In non-competitive inhibition , which a special case of mixed inhibition the lines on double reciprocal
plot converse at one point on X axis which indicates the constant value of Km but the value of Vmax decreases
by increasing inhibitor concentration .
Q5. Abzymes are

[IISc, 2011]

a. enzymes that are highly specific like antibodies

b. antibodies that have catalytic activities
c. also referred to as zymogens
d. enzymes that hydrolyze antibodies
Solution: b. antibodies that have catalytic activities

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Questions from Previous Exams

Questions from Previous Exams

Questions from Previous Exams

352 Concise Chemistry

High Yielding Facts

The mechanism of partially competitive inhibition is similar to that of non-competitive, except that the EIS
complex has catalytic activity, which may be lower or even higher (partially competitive activation) than that of
the enzymesubstrate (ES) complex. This inhibition typically displays a lower Vmax, but an unaffected Km value.
Uncompetitive inhibition occurs when the inhibitor binds only to the enzymesubstrate complex, not to the free
enzyme; the EIS complex is catalytically inactive. This mode of inhibition is rare and causes a decrease in both
Vmax and the Km value.
Product inhibition is often a regulatory feature in metabolism and can be a form of negative feedback
Slow-tight inhibition occurs when the initial enzymeinhibitor complex EI undergoes isomerisation to a second
more tightly held complex, EI*, but the overall inhibition process is reversible. This manifests itself as slowly
increasing enzyme inhibition. Under these conditions, traditional MichaelisMenten kinetics give a false value
for Ki, which is timedependent. The true value of Ki can be obtained through more complex analysis of the on
(kon) and off (koff) rate constants for inhibitor association.
Paclitaxel (taxol), an organic molecule found in the Pacific yew tree, binds tightly to tubulin dimers and inhibits
their assembly into microtubules in the cytoskeleton.
An example of a neurotoxin are the Glycoalkaloids, from the plant species in the Solanaceae family (includes
potato, tomato and eggplant), that are Acetylcholinesterase inhibitors.
Neurotoxicity can also result from the inhibition of receptors; for example, atropine from deadly nightshade
(Atropa belladonna) that functions as a competitive antagonist of the muscarinic acetylcholine receptors
Reversible competitive inhibitors, such as edrophonium, physostigmine, and neostigmine, are used in the
treatment of myasthenia gravis and in anaesthesia.
The organophosphate pesticides such as Malathion, parathion, and chlorpyrifos irreversibly inhibit
acetylcholinesterase.
The herbicide glyphosate is an inhibitor of 3-phosphoshikimate 1-carboxyvinyltransferase, other herbicides,
such as the sulfonylureas inhibit the enzyme acetolactate synthase. Both these enzymes are needed for plants
to make branched-chain amino acids

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