Enzymes: Mechanism of

Action
Importance
Enzymes play an important role in Metabolism:
◦ breakdown of nutrients to supply energy
◦ chemical building blocks; the assembly of those
building blocks into proteins, DNA, membranes, cells,
and tissues
◦ Harnessing of energy to power cell motility and
muscle contraction
◦ Diagnosis: level of enzyme in blood are of diagnostic
importance e.g. it is a good indicator in disease such
as myocardial infarction, liver function tests (LFT) etc
◦ Enzyme can be used therapeutically such as digestive
enzymes.
Define enzymes (Enzymes as Biological Catalysts)
▪Enzymes are proteins that act as catalysts, which are
compounds that increase the rate of chemical
reactions
▪Enzyme catalysts bind reactants (substrates), convert
them to products, and release the products.
▪Although enzymes may be modified during their
participation in this reaction sequence, they return to
their original form at the end.
▪In addition to increasing the speed of reactions,
enzymes provide a means for regulating the rate of
metabolic pathways in the body.
Active Catalytic Sites.
▪The substrate-binding sites overlap in the active
catalytic site of the enzyme, the region of the enzyme
where the reaction occurs.
▪Within the catalytic site, functional groups provided by
coenzymes, tightly bound metals, and, of course, amino
acid residues of the enzyme, participate in catalysis.
Activation Energy and the Transition State.
▪The functional groups in the catalytic site of the
enzyme activate the substrate and decrease the energy
needed to form the high-energy intermediate stage of
the reaction known as the transition-state complex.
 APOENZYME and HOLOENZYME

▪The enzyme without its non protein moiety is termed

as apoenzyme and it is inactive.
▪Holoenzyme is an active enzyme with its non protein
component.
Cofactor
◦ A cofactor is a non-protein chemical compound that
is bound (either tightly or loosely) to an enzyme and
is required for catalysis.
◦ Types of Cofactors:
◦ Coenzymes.
◦ Prosthetic groups.
Types of Cofactors:
1. Coenzyme
▪The non-protein component is loosely bound to
apoenzyme by a non-covalent bond. Examples : vitamins
or compounds derived from vitamins.
2. Prosthetic group
The non-protein component, tightly bound to the
apoenzyme by covalent bonds is called a
Prosthetic group.
Enzyme Specificity

▪Enzymes have varying degrees of specificity for

substrates
▪Enzymes may recognize and catalyze:
- a single substrate
- a group of similar substrates
- a particular type of bond
Activation energy

▪All chemical reactions require some amount of

energy to get them started.
▪It is First push to start reaction. This energy is called
activation energy.
Naming of enzymes

▪The common names for most enzymes derive from

their most distinctive characteristic: their ability to
catalyze a specific chemical reaction. In general,
▪An enzyme’s name consists of a term that identifies
the type of reaction catalyzed followed by the suffix -
ase.
▪For example, dehydrogenases remove hydrogen
atoms, proteases hydrolyze proteins, and isomerases
catalyze rearrangements in configuration.

17
•Sometime the name of the specific substrate
involved is used e.g. xanthine oxidase.

•And also the source of the enzyme e. g. pancreatic

ribonuclease

•Or even the specify its mode of regulation e.g

hormone-sensitive lipase.

•or name a distinguishing characteristic of its

mechanism e. g. a cysteine protease.
• However, to address the ambiguity and confusion
arising from these inconsistencies in nomenclature
and the continuing discovery of new enzymes,
• the International Union of Biochemists (IUB)
developed a complex but unambiguous system of
enzyme nomenclature.
• Enzymes are classified into six functional classes
(ec number classification) by the international
union of biochemists (i.u.b.) on the basis of the
types of
reactions that they catalyze
EC 1. Oxidoreductases EC 4. Lyases
EC 2. Transferases EC 5. Isomerases
EC 3. Hydrolases EC 6. Ligases
 Principle of the international
classification

Each enzyme has classification number

consisting of four digits:
Example, EC: (2.7.1.1) HEXOKINASE
EC: (2.7.1.1) these components indicate the following
groups of enzymes:

2. IS CLASS (TRANSFERASE)

7. IS SUBCLASS (TRANSFER OF PHOSPHATE)

1. IS SUB-SUB CLASS (ALCOHOL IS PHOSPHATE

ACCEPTOR)
1. SPECIFIC NAME
ATP,D-HEXOSE-6-PHOSPHOTRANSFERASE (Hexokinase)
 6 CH 2OH 6 CH OPO 2−
2 3
ATP ADP
5 O 5 O
H H H H
H H
4 1 4 H 1
OH H OH
Mg2+
OH OH OH OH
3 2 3 2
H OH Hexokinase H OH
glucose glucose-6-phosphate

1. Hexokinase catalyzes:
Glucose + ATP → glucose-6-P + ADP
Oxidoreductases, Transferases and Hydrolases
Lyases, Isomerases and Ligases
Mechanism of Action of Enzymes
1. Enzymes increase reaction rates by
decreasing the Activation energy
2. Enzyme-Substrate Interactions: Lock-
and-Key Model
3. Enzyme-Substrate Interactions: Induced
Fit Model
4. Enzyme-Substrate Interactions:
Formation of Enzyme substrate
1. Enzymes increase reaction rates by decreasing the Activation energy

Enzymes
Lower a
Reaction’s
Activation
Energy
 2. Lock-and-Key Model
In the lock-and-key model of enzyme action:
- the active site has a rigid shape
- only substrates with the matching shape can fit
- the substrate is a key that fits the lock of the active site
This is an older model, however, and does not work for all
enzymes
3. Induced Fit Model
In the induced-fit model of enzyme action:
- the active site is flexible, not rigid
- the shapes of the enzyme, active site, and substrate adjust to
maximumize the fit, which improves catalysis
- there is a greater range of substrate specificity
This model is more consistent with a wider range of enzymes
 Enzyme-substrate complex
Step 1: Enzyme and substrate combine to form complex

E + S ES
Enzyme Substrate Complex

+
 Enzyme-product complex

•Step 2: An enzyme-product complex is formed.

ES EP

ES transition EP
state
 Product
•The enzyme and product separate

•EP E + P
The product
is made
Enzyme is
ready
EP for
another
substrate.
ENZYMES EMPLOY MULTIPLE MECHANISMS TO
FACILITATE CATALYSIS
Four general mechanisms to achieve dramatic
catalytic enhancement of the rates of chemical
reactions.
1. Catalysis by Proximity
For molecules to react, they must come within
bond forming distance of one another. The higher
their concentration, the more frequently they will
encounter one another and the greater will be
the rate of their reaction.
2. Acid-Base Catalysis
The ionizable functional groups of aminoacyl side
chains and (where present) of prosthetic groups
contribute to catalysis by acting as acids or bases.
3. Catalysis by Strain
Enzymes that catalyze lytic reactions which involve
breaking a covalent bond typically bind their substrates
in a conformation slightly unfavorable for the bond
that will undergo cleavage. The resulting strain
stretches or distorts the targeted bond, weakening it
and making it more vulnerable to cleavage.
4. Covalent Catalysis
The process of covalent catalysis involves the formation
of a covalent bond between the enzyme and one or
more substrates. The modified enzyme then becomes a
reactant. Covalent catalysis introduces a new reaction
pathway that is energetically more favorable—and
therefore faster—than the reaction pathway in
homogeneous solution.
Examples
What Affects Enzyme Activity?

•Three factors:
1. Environmental Conditions

2. Cofactors and Coenzymes

3. Enzyme Inhibitors

51
1. Environmental Conditions
1. Extreme Temperature are the most
dangerous
- high temps may denature (unfold) the
enzyme.
2. pH (most like 6 - 8 pH near neutral)
3. substrate concentration .

52
2. Cofactors and Coenzymes
•Inorganic substances (zinc, iron) and vitamins
(respectively) are sometimes need for proper
enzymatic activity.

•Example:
Iron must be present in the quaternary
structure - hemoglobin in order for it to pick
up oxygen.

53
Environmental factors
Optimum temperature The temp at which enzymatic reaction occur fastest.
Environmental factors
pH also affects the rate of enzyme-substrate
complexes
◦ Most enzymes have an optimum pH of
around 7 (neutral)
◦ However, some prefer acidic or basic
conditions
Substrate Concentration and Reaction Rate
The rate of reaction increases as substrate
concentration increases (at constant enzyme
concentration)
Maximum activity occurs when the enzyme is
saturated (when all enzymes are binding substrate)
Enzyme Inhibitors
Competive - mimic substrate, may block active site, but may dislodge it.
Enzyme Inhibitors
Noncompetitive
Enzyme Kinetics
Enzyme kinetics
▪The field of biochemistry that deals with Quantitative
measurement of the rates of enzyme catalyzed
reactions.
▪And also the systematic study of factors that affect
these rates.
▪Kinetic analyses allows reconstruction of the number
and order of the individual steps by which enzymes
transform substrates into products.
▪The study of enzyme kinetics helps in the identification
of potential therapeutic agents that selectively enhance
or inhibit the rates of specific enzyme-catalyzed
processes.
Enzyme kinetics
Together with site directed mutagenesis and other
techniques that probe protein structure, kinetic
analysis can also reveal details of the catalytic
mechanism.
A complete, balanced set of enzyme activities is of
fundamental importance for maintaining
homeostasis.
An understanding of enzyme kinetics thus is
important for understanding how physiologic
stresses such as anoxia, metabolic acidosis or
alkalosis, toxins, and pharmacologic agents affect
that balance.
And also the systematic study of factors that affect
these rates
Enzyme kinetics began in 1902 when Adrina Brown
reported an investigation of the rate of hydrolysis of
sucrose as catalyzed by the yeast enzyme inveratase
According to Brown, when substrate (sucrose)
concentration is much higher than that of the enzyme,
reaction rate becomes independent of sucrose
concentration
Thus, overall reaction is composed of two elementary
reactions in which the substrate forms a complex with
the enzyme that subsequently decomposes to products
and enzymes

Where E, S, ES and P symbolize the enzyme, substrate,

enzyme-substrate complex and products, respectively
The overall rate of production of [ES] – Difference between the
rates of elementary reactions leading to its appearance and
those resulting in its disappearance.

At this point, an assumption is required to achieve an analytical

solution:
◦ The rapid equilibrium assumption: Michaelis - Menten
Approach.
◦ The steady-state assumption: Briggs and Haldane Approach.
 Michaelis – Menten Equation
For the enzyme catalyzed reaction: assumes a rapid equilibrium
between the enzyme and substrate to form an [ES] complex.

The equilibrium constant Km can be expressed by the following

equation in a dilute system.
Since the enzyme is not consumed, the conservation equation
on the enzyme yields.

Then rearrange the equilibrium constant equation.

Substituting [E] in the above equation with enzyme mass

conservation equation
Then the rate of production formation v can be expressed in terms
of [S]

Where
Steady State Assumption (SSA)
At a steady state synthesis of
ES must equals to its
consumption over the course
of reaction i.e. ES maintain
steady state

Transition phase of the

reaction [ES] remains constant
until the substrate is nearly
exhausted.
SSA and Rate Equation

Now: Base on steady state assumption, d[ES]/dt = 0

d[ES]/dt = k1[E][S] –k-1[ES] – k2[ES] = 0
(steady state assumption)
solve for [ES] (do some algebra)
[ES] = [E][S] k1/(k-1 + k2)
Define KM (Michealis Constant)
KM = (k-1 + k2)/k1 => [ES] = [E][S]/KM
 SSA lead to Michaelis - Menten
Then the rate of production formation v can be expressed in
terms of [S]

Where

Michaelis Menten Equation

 Km
KM is the substrate concentration
required to reach half-maximal
velocity (vmax/2).

KM is a measure of a substrate’s
affinity for the enzyme.

A small Km means the substrate

binds tightly to the enzyme and
saturates the enzyme
 Vmax
Considering the total enzyme concentration the maximal
rate, that the enzyme can attain is Vmax.
Vmax is equal to the product of the catalytic rate constant
(kcat) and the concentration of the enzyme.
The Michaelis-Menten equation can then be rewritten as V=
Kcat [Enzyme] [S] / (Km + [S]).
Kcat is equal to K2, and it measures the number of substrate
molecules "turned over" by enzyme per second.
The higher the Kcat is, the more substrates get turned over
in one second.
Lineweaver-Burk Equation

Starting with the MM equation

Reciprocal of MM equation

Lineweaver-Burk Equation

Equation is the equation for a straight line,

y = ax + b, where y = 1/v0 and x = 1/[S].
Lineweaver-Burk Equation
A plot of 1/v0 as y as a function of
1/[S] as x therefore gives a straight
line whose y intercept is 1/Vmax and
whose slope is Km/Vmax.

Such a plot is called a double

reciprocal or Lineweaver-Burk plot

Setting the y term of equation equal

to zero and solving for x reveals that
the x intercept is −1/Km
Lineweaver-Burk plot, has the great advantage of allowing a
more accurate determination of Vmax, which can only be
approximated from a simple plot of V0 versus [S].

The double-reciprocal plot of enzyme reaction rates is very

useful in distinguishing between certain types of enzymatic
reaction mechanisms.
Kinetics of Isosteric enzymes
Isosteric enzymes
(with only one
enzyme
conformation, 1), the
efficiency of substrate
binding (dashed
curve) declines
constantly with
increasing [A],
because the number
of free binding sites is
constantly
decreasing.
Kinetics of allosteric enzymes
Allosteric enzymes, the
binding efficiency initially
rises with increasing [A],
because the free enzyme is
present in a low-affinity
conformation (square
symbols), which is gradually
converted into a higher-
affinity form(round symbols)
as a result of binding with A.
It is only at high [A] values
that a lack of free binding
sites becomes noticeable
and the binding strength
decreases again.
Enzyme regulation
 Inliving systems hundreds of different
enzyme catalysed reactions occur
simultaneously.

 These reactions must be regulated for the

proper functioning of a living system.

 Regulatory enzymes exhibit increased or

decreased catalytic activity in response to
certain signals. 3
An enzyme’s catalytic activity can be
directly controlled through structural
alterations that influence the enzyme’s
substrate-binding affinity.
• Allosteric Enzyme Regulation
• Proteolytic Activation of enzymes
• Reversible Covalent Modifications
• Regulation by Isoenzymes
4
❑ Enzymatic activity can be activated or inhibited
through non-covalent interaction of the enzyme
with metabolites other than the substrate. This form of
control is termed Allosteric regulation.

❑ Allosteric proteins contain distinct regulatory sites

and multiple functional sites.

5
 Many of the ideas about ligand-
induced conformational changes
of enzymes developed as a result
of work on the biosynthetic
pathways of microorganisms.

 In 1950s, it was found that

Threonine dehydratase, the first
enzyme in the Isoleucine
biosynthesis pathway was strongly
inhibited by the end-product
Isoleucine. 6
 Feedback inhibition: The committed step in a
biosynthetic pathway is inhibited by the ultimate end
product of the pathway.

 The feedback inhibitor F bears little structural similarity

to A, the substrate for the regulatory enzyme E1.

 F acts on a binding site distinct from the substrate

binding site. 7
Activation and inhibition
Allosteric proteins show the property of cooperativity i.e.,
activity at one functional site affects the activity at others. A
slight change in substrate concentration can produce
substantial changes in activity.

Their kinetics do not obey the

Michaelis–Menten equation.
Their V versus [S] plots yield
sigmoid curves rather than
hyperbolas.

9
 Positive cooperativity: Ligand binding at one
site facilitates the binding of other sites on the
same molecule.

 Negative cooperativity: Ligand binding at

one site inhibits the binding of other sites on
the same molecule.

10
 Regulatory enzymes for which substrate and
modulators are identical are called
Homotropic.
 When the modulator is a molecule other
than the substrate, the enzyme is said to be
Heterotropic.

 Regulatory enzymes are also subject to an

activation process by a metabolite which
belongs to another metabolic pathway, which
11
 Allosteric
enzymes typically have an
oligomeric organization.

 They are composed of more than one

polypeptide chain (subunit), and each subunit
has a binding site for substrate, as well as a
distinct binding site for allosteric effectors.

 The regulatory effects exerted on the enzyme’s

activity are achieved by conformational 12
 Glycogen phosphorylase is dimer of two identical
subunits.
 ATP and Glucose-6-phosphate are negative
heterotropic effectors. ATP is a feedback inhibitor.
 AMP is a positive heterotropic effector (activator).

17
Isoenzymes are enzymes that differ in amino
acid sequence yet catalyze the same reaction.

18
 Isoenzymes are enzymes that differ in amino acid
sequence yet catalyze the same reaction.
 These enzymes display different kinetic parameters,
such as Km, or different regulatory properties.
 Encoded by different genetic loci, (arise through
gene duplication and divergence).

 Allozymes - Enzymes that arise from allelic variation

at one gene locus.

 The existence of isozymes permits the fine-tuning of

metabolism to meet the particular needs of a given 19
 Lactate dehydrogenase catalyses the reaction:

 In the reverse direction, it represents the last step in the

anaerobic glycolysis for the regeneration of NAD+
required for G3PDH reaction.
20
 The functional enzyme is a tetramer and there are 5 forms
of the enzyme.

 Human beings have two isozymic polypeptide chains for

LDH: the H isozyme highly expressed in heart and the M
isozyme found in skeletalmuscle.

21
Zymogens are inactive precursors of enzymes.

Zymogens or proenzymes acquire full activity only upon

specific proteolytic cleavage of one or several of their peptide
bond.

Irreversible process.

22
• Some protein hormones
are synthesized in the
form of inactive precursor
molecules, from which the
active hormone is derived by
proteolysis.

Insulin, an important
• metabolic regulator, is
generated by proteolytic
excision of a specific
23
 The digestive enzymes are synthesised as zymogens
in the pancreatic acinar calls and stored as zymogen
granules.

 Enteropeptidase catalyses the activation of

trypsinogen as it enters the duodenum.

 Trypsin catalyses the activation of other zymogens.

 Trypsin action involves the peptide bond at the C-

terminal side of lysine or arginine side chain. 25
 Premature activation of zymogens is
prevented to reduce damage of the pancreas :
▪ by the presence of Trypsin inhibitor protein in
the pancreatic secretion
▪ initial trigger by enteropeptidase at a site distinct
from the site of production of zymogens.

26
 •
Activ tion of 0 Sill

Chymoirypsinogcn (inactive
zyrnogcn) I i

Cleavage al Atg15
by trypsin

1r-Chymotrypsin (active enzyme)

i I

Self-digestion al Leu
13, 146, and Asn
Tyr by
1r<hymotrypsi148n

Ser Arg Tiu Asn

o-Chymctrypsin (active enzyme)

Leu De Tyr AJ
IS I a

27
 The formation of blood clots - series of zymogen
activations.

 Thrombin (a serine protease) specifically cleaves

Arg–Gly peptide bonds of fibrinogen and convert it
into fibrin.

 Fibrin readily aggregates into ordered fibrous

arrays that are subsequently stabilized by covalent
crosslinks.
28
29
The covalent attachment of a molecule can modify the
activity of enzymes and many other proteins.

A donor molecule provides a functional moiety that

modifies the properties of the enzyme.

Most modifications are reversible.

30
Cova ent Modification
Table 10.1. Common covalent of protein
modifications activity

Modificati Donor Example ofmodified protein Protein

on molecule function
Phosphorylat AT Glycogen Glucose homeostasis;
ion P phosphorylase energy
Acetylation Acetyl CoA Histones transduction
Myristoylatio Myristoyl CoA DNA packing;
Src
n transcription
NAD RNA
Signal transduction
ADP- Farnesyl pyrophosphate polymerase
ribosylation Transcription
HC03 - Ras
Farnesylation Signal transduction
Thrombin
31-Phosphoadenosiue-51- Blood clotting
y.Carboxylati
phosphosulfate Fibrinogen
on Blood-clot formation
Ubiquitin Cyclin
Sulfation Control of cell cycle
Ubiquitinatio
n
Phosphorylation and dephosphorylation are the most common
31
 Some proteins such as Ras and Src are
localized to the cytoplasmic face of the plasma
membrane by the irreversible attachment of a
lipid group.

 The attachment of ubiquitin is a signal that a

protein is to be destroyed.

 Cyclins must be ubiquitinated and destroyed

before a cell can enter anaphase and proceed 32
 The transfer of a phosphate group from a donor to
an acceptor amino acid of a protein.
 Phosphorylation : by Kinases
 Phosphatases :remove the phosphate group
through hydrolysis of the sidechain phosphoester
bond.

33
 Glycogen phosphorylase, the enzyme that
catalyses the release of glucose units from
glycogen.
 Regulated by both phosphorylation and
allosteric regulation.

34
 I s la
• Muscle glycogen phosphorylase ~ , ,_<~_val
,e_n_1_c_on_~_o_l , ,::>
is a dimer of two identical Phosphory law kinase

subunits. Phosphoprotei phosphatase 1

n

• Each subunit contains an active Pho phorylase

/1
Phospborylase
a
I naciive Inactive
(T tare
site and an allosteric effector site (T state) )

near the subunit interface. AM ATP 0

P .! Glucos
e
::
Clucose- 0 e
6-P
Glucose
-
I.I

c,
-
c Caffeine

Caffeine 0
~Ia
I .
c
.. I I z0
.

Phosphorylase b Phosphorvlase
Active a
(R state) Active
(R state) 35
Covalent modification through
phosphorylation of Ser14 in
glycogen phosphorylase converts
the enzyme from a less active,
allosterically regulated form (the b
form) to a more active,
allosterically unresponsive form
(the a form).

36
 The reaction of ADP-ribosylation is catalysed by specific
enzymes, ADP-ribosyl transferases, which use NAD+ as a
substrate.

 In humans, one type of ADP-ribosyltransferases are the NAD+:

arginine ADP-ribosyltransfe rases, which modify amino acid
residues in proteins such as histones by adding a single ADP-
ribose group.

37
38
Significance – In
EukAacrtiyvaotteedsby DNA cleavages –involved in
DNA repair, transformation and cellular differentiation.

 Modifies histones – causes changes in chromatin

structure.

 ADP-ribosylation modulates the activity DNA ligase

III, terminal deoxynucleotidyl transferase, α and β
DNA polymerases, topoisomerases, Ca++ and Mg++
dependant endonucleases.
39
 The activity of ADP-ribosyl transferase is significantly
 The ADP-ribosylation reactions play a role in the
toxicity of certain bacteria.

 Diphtheria toxin inhibits the EF2 elongation factor

by mono-ADP-ribosylation, which blocks protein
synthesis in the infected cell.

 Choleric and pertussis toxins provoke ADP-

ribosylation of a G protein and causes regulation of
adenylate cyclase activity, lead to increase in the
cellular ratio of cyclic AMP.
40
 Protein glycosylation is a post-translational
modification.

 There are two major types of glycosylation, N- and O-

glycosylations, which involve the binding of the
saccharide chain to an asparagine and a serine or a
threonine, respectively.

 N-glycosylations occur on an asparagine belonging to a

sequence Asn-X-Ser/Thr, where X can be any amino
acid except proline or aspartic acid.

 O-glycosylations occur on hydroxyl groups of a serine 41

2nd step in the formation of typical O-linked oligosaccharides
in proteins such as glycophorin : A specific glycosyltransferase
catalyzes addition of a galactose residue from UDP-galactose
to C3 of N-acetylgalactosamine attached to a protein forming a
β13 linkage.

42
 This type of covalent modification consists of the
binding of an adenyl group to a well-defined tyrosine
residue of a protein.
 Eg: Glutamine
Synthase
 Glutamine synthetase (GS) catalyses the ATP-
dependent condensation of to ammonia with glutamate,
yield glutamine.

 Bacterial GS regulated by adenylation. Mammalian

and plant GS not regulated by adenylylation.
43
Adenylation of glutamine
synthetase, catalyzed by the
enzyme adenylyl transferase
(ATase)
Involves the phosphodiester
bond between the OH group
of the Tyr in glutamine
synthetase and the phosphate
group of an AMP nucleotide.

44
 DNA ligases and RNA ligases catalyze the
formation of phosphodiester bonds at single
strand breaks with adjacent 3’ hydroxyl and 5’
phosphate termini in DNA or RNA,
respectively.

 The first step in the catalytic cycle is the

adenylylation of an active-site ε-NH3 group of
Lys residue.
45
Adenylation

E-(Lys)-NH2

0 0 0 H O
II/\ II II I II
Ad-O-P-O-P-0-P-o- == E-(Lys)-N+-p-
I I I I I
o- o- o- H o-

H O
I II
E-(Lys)-N-P-0-Ad +
I I
H o-

46
• Rapid change in the amount of active enzymes.
• Large amplification of the initial signal

• System is always poised for activation or inactivation.

• System can be rapidly activated, since it is reversible in nature.
• When stimulus removed, system rapidly converted back to resting
state.
47
As enzymes are protein in nature, they are synthesized
from amino acids under gene control and degraded
back to amino acids after its action.

Enzyme quantity depends on the rate of enzyme

synthesis and the rate of its degradation.

48
 Cellularphysiology and differentiation
processes require that at well defined stages
the enzyme activity should start and end.

 The process of enzyme turnover necessary for

the cell to adapt to changes in the environment
and to remove an abnormal enzyme.

 The process of peptide bond hydrolysis in

enzymes do not liberate energy, unlike peptide
bond formation which utilises ATP. 49
 Intracellular protein degradation occur both
lysosomally and proteasomally.
 The process may be energy dependent or
independent.
 It can be selective or non selective
degradation.
 Many house keeping enzymes are degraded by
lysosomal degradation.
 The short lived enzymes are degraded by extra-
lysosomal mechanisms and are present in 50
 Proteasomes are large multisubunit proteases that
degrade ubiquitin-tagged proteins in an ATP
dependent manner.
 Ubiquitin is a small basic peptide of 76AA in
found eukaryotic cells.
 Proteasomes degrade proteins into short peptides that
ar e rapidly hydrolysed by cytoplasmic exopeptidase s.

51
 Lysosomesperform 2 processes: Phagocytosis
and Autophagy.

 Lysosomes contain different hydrolytic

enzymes which have their optimum pH at
acidic range.

 Enzymes degraded by this way do not require

ubiquitination.
52
 Fundamentals of Enzymology: The Cell and
Molecular Biology of Catalytic proteins, 3rd
Edition – Nicholas. C. Price, Lewis Stevens.
 Enzymes : Biochemistry, Biotechnology, Clinical
Chemistry – Trevor Palmer.
 Enzyme catalysis and Regulation – Gordon G.
Hammes.
 Biochemistry, 5th edition – Jeremy.M.Berg, John
L. Tymoczko, Lubert Stryer.
 Biochemistry, 6th edition – Reginald.H.Garrett,
Charles. M Grisham. 54