Introduction to Biochemistry

1
INTRODUCTION TO BIOCHEMISTRY (4
hrs. )
- Introduction:
• Definitions
• Role of biochemistry
- Cellular components
- Brief introduction to metabolism &
Enzyme
References
Lehninger Principles of Biochemistry
(Page No. 3-12)

2
Learning
outcomes
— Define medical biochemistry
— Describe historical development of
biochemistry
— Explain the scope of biochemistry
— Explain the role of biochemistry in health
science
— List and describe cellular components
— Explain metabolism & the role of enzymes in
metabolism
3
 Introduction
Definition of Biochemistry
• Biochemistry is literally the chemistry of life

• The science concerned with the various molecules

that occur in living cells and organisms and with
their chemical reaction

• “Biochemistry is the study of carbon compounds that

crawl”,
Mike Adams

• “We think we have found the basic mechanism by 4

 Cont…
• Biochemistry can be defined as the science concerned
with the
chemical basis of life (Gk bios “life”).

• The cell is the structural unit of living systems and

biochemistry can also be described as the science
concerned with the chemical constituents of living cells
and with the reactions and processes they undergo

• Anything more than a superficial comprehension of life

– in all its diverse manifestation - demands a
knowledge of biochemistry 5
Cont…

• Biochemistry describes living organisms in

molecular terms :
structures
mechanisms
chemical processes shared by all organisms
 organizing principles that underlie life
in all its diverse forms
function
• *the molecular logic of life”

6
 Cont…
• Therefore, biochemistry:
 is the application of chemistry to the
study of biological process at cellular & molecular
level

 aimed to describe & explain, in

molecular terms, all chemical processes of
living cells

 biochemists have isolated numerous molecules

found in cells, determine their structures and
analyze how they function
7
 Historical Development of
• Biochemistry
Justus von Liebig introduced the concept of
metabolism, 1842

• Biochemistry has developed as an offshoot of organic

chemistry "physiological chemistry"

• Biochemistry" was coined by Neuberg,1903 , Gks,

bios= life & chymos = juice

• It emerged as a distinct discipline around the

beginning of the 20th
century

• When scientists combined chemistry, physiology and

biology to investigate the chemistry of living 8
systems
Cont.….
Two important discoveries

• Discovery of the role of enzymes as catalysts

 Büchner (1899) experiment: yeast cells extract could

catalyze
the fermentation of glucose into alcohol & CO2

 Jems sumner showed that enzymes are proteins

• Identification of nucleic acids as information molecules

 Flow of information from nucleic acids to

9
proteins DNA →→ RNA →→ Protein
 Transgenic
Gene engineering animal

HGP from
Clone 1990, 1
completed in 0
Some milestones events of
Biochemistry

11
Cont…
• The Nobel Prize in Medicine 2017
 “For their discoveries of molecular mechanisms
controlling the
circadian rhythm"

Jeffrey C. Hall Michael W. Young

Michael Rosbash

1
2
Scope of
Biochemistry
• The scope of biochemistry is as vast as life itself !
 Every aspect of life-birth, growth, reproduction, aging,
disease…
• Because life depends on biochemical reactions,
biochemistry has become the basic language of all life
sciences

• Be essential to all life sciences as the common knowledge

 Genetics,
 Cell Biology
 Molecular Biology
 Physiology
 Immunology
 Pharmacology
 Toxicology
 Pathology
 Microbiology
 Nutrition 13
Cont.….

•Biochemistry is a contemporary
science
•Has three main focuses
─Metabolism
─Structural Biochemistry
─Molecular Genetics
• Other branches of biochemistry
– Endocrinology – Toxicology – Bioinformatics
– Immunology – Oncology – Research
– Enzymology – Biotechnology biochemistry
– Environmental – Industrial – Biotechnology
biochemistry biochemistry – Clinical
– Food & nutrition – Agricultural biochemistry
biochemistry – Medical 14
biochemistry
Role of biochemistry
• Why study Biochemistry?
1. Biochemical studies lead us to a
fundamental understanding of life
How does the body works
 Biochemical similarities and
differences among many forms of life
 How do organisms store &
transfer information to perpetuate
generation
Molecules and processes
Production of cellular energy 15
Cont…
2. Biochemistry has a profound impact in understanding
of medicine, health, nutrition, and the
environment

• Biochemical study results have led to molecular

understanding of diseases (diabetes, cancer, sickle-cell
anemia, phenylketonuria, hypercholesterolemia,
…….)
•
• Human genome sequencing helps to find cures for
disease (Alzheimer's disease, depression,
influenza, other disease)

• Recombinant DNA technology will play a major 16

role in the diagnosis and treatment of diseases
 Cont…
• The study of enzymes and metabolism provides a
foundation for the rational design of drugs and for the
detailed understanding of nutrition

• Biochemistry used as tools for research in all branches

of medical science:
 Studying molecular & cellular mechanism of
disease to cure different untreated disease such as
- Cancer
- Obesity Future Hope
- Diabetic
- Neurodegenerative 1
7
diseases
active areas of biochemical research

Active areas
of
biochemical
research

18
Cont…
• Biochemistry & medicine enjoy a mutually
cooperative relationship
The two major concerns for health sciences workers:
– the understanding & maintenance of health
– the understanding & effective treatment of diseases
 Biochemistry impacts enormously on both of these
fundamental concerns of medicine
 Health depends on a harmonious balance of
biochemical reactions occurring in the body
 Disease reflects abnormalities in biomolecules &
biochemical reactions (electrolyte imbalance, defective
nutrient, hormonal imbalances, genetic disorders) 1
9
Cont…
• The interrelationship of biochemistry and medicine is a
wide, two- way street

Examples of the two-way street connecting biochemistry 2

and medicine
 Cont.…
Treatment
Diagnosis

Biochemistry

Prevention
21
Cellular components
• Cells are highly organized & requires constant
source of energy to maintain the ordered state

• All cells have similar basic structure

Consists of :
Non-protoplasmic components

Protoplasmic components

22
 Non-protoplasmic
components
• All cell parts are made of highly organized groups of
few types of biomolecules:
 Nucleic acid and protein……..17%
 Lipid……………..13.8%
 Carbohydrate………….1.5%
 H2O…………..60%
 Minerals & vitamins…6.1%
 Other……… trace

23
Cont.…
• Chemical foundations

•21 of naturally occurring

elements are commonly
presented in living cell

•99% of its mass accounts to

O,N,H & C

•Inorganic ions (Na,K,Ca,Mg,Ca,Cl)

make about 0.5%

•The remaining elements are

24
referred as
Major types of functional groups found in biochemical
compounds of the
human body

25
26
Cont.…
•The chemistry of living organisms is organized
around carbon

2
8
• Macromolecules are the major constituents
of cells
 Proteins
 Polysaccharides
 Lipids
 Nucleic acids

29
 Cot…
• Which came first—
 catalysts or
hereditary molecules?

• Originally,
 RNA was catalyzing and
encoding its own replication

30
Protoplasmic components
•There are three distinct domains
of life

31
 Cont…
• All cells of the simplest and most complex
organisms share certain fundamental properties

↳
Cytoplasm ⟹ cytosol & suspended
Plasma membrane

↳ Prokaryotes ↳ Eukaryoteso esCoenzyme

particles o Organelle
 Nucleoid or s
nucleus o Metabolit

? ? s

3
Cont….
•Cell membrane:
 Defines periphery of the cell
 Composed of mainly lipid &
proteins
 Transports ions and molecules in
& out
 Receptors transmit signal into
cells
 Membrane enzyme
participate in reaction
 Allows change in shape and size 33
Cont….
•Chemically Cell membrane is composed
of:
 Lipid
 Protein
 Carbohydrates
 For instance human RBC contains
protein (57%),lipid (42),
• The most widely accepted models
of cell membrane is fluid mosaic
model
34
Cont…

Cytoplasm
• Cytosol with variety of suspended
particles with specific functions
 Enzymes
 Metabolites
 Coenzenzymes…

• All organelles
 Mitochondoria
 ER….

35
Cell…
• Eukaryotic cells contain various organized
structures

• Cell organelles

Membranous
Mitochondriaorganelles  Ribosomes organ
Non-membranous
 Endoplasmic  Cytoskeleton
- microtubules,
reticulum - actin filaments
 Golgi complexes - intermediate
 Peroxisomes filaments
 Lysosomes  Centrioles
 Vacuoles  Proteasomes
 Chloroplasts
36
Cont.…
Mitochondria
• Is site for:
 Aerobic ATP
synthesis
 Ketone body
synthesis
 Amino acid
metabolism
 Glucose metabolism
 Heme synthesis
 Steroid hormone
synthesis……..
37
38
Cont.…
Endoplasmic reticulum & Golgi
complexes
• Site for the synthesis and
processing of lipids and
membrane proteins

Ribosom
e

39
 Cont…

Lysosomes
• Suicide bags’ of the cell, which are found
only in
animal cells

• ATP-driven proton pumps accumulate H+

in the
lysosomes w/c make pH values of 4.5–5

• Intracellular organelles of digestion

containing different types of
hydrolases

• Break down macromolecule ,worn 4

0
out cell organelles, cellular debris
41
 Cont…

Peroxisomes
• probably formed by budding SER

• carryout oxidation reactions in which

hydrogen peroxide (H2O2) is produced

• Peroxisomes protect cells from the

potentially damaging effects of free
radicals

• Site for oxidation of very long chain fatty

acids (≥20 C), conversion of cholesterol to
bile acids & the synthesis of ether lipids
• Catalase and peroxidase destroys the
(plasmalogens)
peroxides &
other free radicals
4
Cont.…
Ribosome
• Found attached to rough ER or
floating free in cytosol

• Produced in a part of the

nucleus,
nucleolus

• Fixed ribosomes manufacture

protein enter the ER (modified &
packaged for secretion)

43
Cont…

44
Cont…
Proteosome
• Barrel shaped complex with
protein degrading enzyme

• Site of protein hydrolysis

• Degrades ubiquitin tagged

protein

45
 Cont…
Cytoskeleton
• complex network of
protein filaments

• Provides shape and

structure

• Helps move organelles

around the cell
• Made of three types of
filaments
Microtubules
46
Microfilaments
Cont…
• Assembly and disassembly of the spindle
structures during mitosis

47
 Cont…
Nucleus
• Most prominent organelle of the
cell
• Found in all human cells, except
matured RBC
• Control & the information of
center (DNA)
• DNA+ Histone = chromatin
• Surrounded by a double
membrane
• Usually one per cell

48
4
9
 Introduction to metabolism & Enzyme
• Metabolism: the chemical reactions that occur in living
organism

• These reactions are catalyzed by enzymes & the reactant

compounds are called metabolite

• Anabolism- energy requiring biosynthetic pathways

• Catabolism- degradation of fuel molecules and the production

of energy for cellular function 50
 Catabolism
• The purpose of catabolism is to trap energy of the
biomolecules in the form of ATP & to generate the
substance required for the synthesis of complex
molecules

• Conversion of complex molecules into their building blocks

• Formation of simple intermediates

51
Cont…
Catabolism

5
 Cont…
Anabolism
• Consume energy to synthesize large
complex molecules, the starting materials
are relatively few.
 pyruvate, acetyl CoA , TCA
intermediates

• Anabolic reactions are dependent on the

supply of
energy (as ATP or GTP)
reducing equivalents (as NADPH +H+)
• The synthesis of carbohydrate is an example
anabolism 5
Anabolism

54
 Metabolic Pathways
• Metabolisms are directed along specific sequences,
metabolic pathways

• Most of metabolic pathway reactions are catalyzed

by specific enzyme

55
 Types of Metabolic Pathways

Catabolic Anabolic Amphibolic

• Degrade large • Synthesis of larger • Both catabolic and
molecule into smaller molecules from smaller anabolic in nature
molecules molecules
• Produce ATP,NADPH • Use ATP,NADPH,
NADH

• e.g. • e.g. • e.g.

Glycolysis,β-oxidation, Gluconeogenesis, fatty acid TCA
amino acid oxidation synthesis, glycogen cycle
synthesis, lipid synthesis,
protein synthesis

56
57
Cont…
• There are many possible biochemical reactions
however they fall into a few types:

⟺ aldehyde
 Oxidation & reduction: e.g. alcohol

 Functional groups transfer: e.g. Pi, -

CH3

 Addition & removal of water: e.g. hydrolysis

 Bond making /breaking reactions: e.g. 5

8
carbon- carbon bond breakage
 Bioenergetics (thermodynamics)
•Definition

 The quantitative study of energy transductions

that occur in living cells

•Changes in free energy (ΔG) provide a measure of the

energetic
feasibility of a chemical reaction

•Bioenergetics predicts possible processibility

•Biological energy transformations obey the

laws of thermodynamics 59
Cont.….
•First law of thermodynamics
 Total energy of a reaction system remains
constant

•Second law of thermodynamics

 Total entropy of a system must
increase if a process is to occur
spontaneously
• At constant temperature and
ΔG ΔH the
=
•Combines - two laws of pressure:
 Gibbs free energy (G)
TΔS
thermodynamics:  ΔH =the change in enthalpy
(heat)
 T = absolute temperature
60
 ΔS= change in entropy
Cont.…
•In biochemical reactions ΔH= ΔE (change in
energy of the
reaction) ΔG= ΔE -
TΔS
• ΔG can be used to predict the direction of a
reaction

 ΔG = negative → loss of free energy

(exergonic) & the reaction goes
spontaneously A to B

 ΔG = positive → gain of free energy

(endergonic) &
energy must be added to run the reaction 61
Exergonic reaction Endergonic reaction
63
Cont.…
•The ΔG of the reaction A → B depends on the
concentration of the reactant and product
• Δ Go = standard free energy change

[𝐀
• R = gas constant (1.987 cal/mo. Degree)
ΔG= ΔG o + RT
[𝐁]
• T = absolute temperature ( oK)

]
• [A] and [B] = actual conc. of the reactant &
ln product
• ln= represents the natural logarithm
• ΔGo is the free energy change, under standard conditions
( T=298 K (250C), conc.= 1M(1mol/L), P= 101.3kPa or 1 atm)

•Under these conditions , the ratio of products to reactants is

zero , and
ln1 = 0
64
Coupling of endergonic & exergonic
processes
•The vital processes obtain energy by
coupling to oxidative reactions in
cells

•The conversion of A to B occurs

with release of free energy

• Free energy is required to

convert metabolite C to
metabolite D

66
 C6H12O 36
6 ATP

ΔG°′ = −2850 ΔG0= 7.3

kJ/mol kJ/mol

6CO2 + 36 ADP + 36
6H2O Pi

Coupling of glucose oxidation & ATP

synthesis

67
The high-energy
compounds
•The complete oxidation of a metabolic fuel
releases free energy ( e.g. glucose &
palmitate)
 C6H12O6 + 6 O2 → 6 CO2 + 6 H2O, ΔG°′ =
−2850 kJ/mol

 C16H32O2 + 23 O2 → 16 CO2 + 16 H2O, ΔG°′ =

−9781 kJ/mol
• Released free energy canbe
UTPrecovered by the
 Creatine
synthesis
phosphate
of a few types of “high-energy”
 intermediates
NADH 68
ΔG° of phosphate hydrolysis of some compounds of common in living
system
Compound ΔG°
kJ · mol−1 kcal· mol−1
Phosphoenolpyruvate −61.9 -14.8
1,3-Bisphosphoglycerate −49.4 -11.8
ATP (→ AMP + PPi) −45.6 -7.7
Phosphocreatine −43.1 -10.3
Acetyl phosphate −42.3 10.1
ATP (→ ADP + Pi) −30.5 -7.3
Glucose-1-phosphate −20.9 -5.0
PPi −19.2 -4.6
Fructose-6-phosphate −13.8 -3.8
Glucose-6-phosphate −13.8 -3.3
Glycerol-3-phosphate −9.2 -2.2
NB. The compounds above ATP can spontaneously transfer a phosphoryl group to ADP
to for m
Cont…
•Low - energy compounds
 ΔGo less than that of ATP
 Glucose-6-P (-4.6 kcal/mole)
 PPi (3.3 kcal/ mole)
•High-energy compounds
 ΔGo higher than that of ATP
 Phosphoenolpyruvate
 creatine phosphate (- 10.3
kcal/mole)

70
71
Cont.…

•ADP can accept high-

energy phosphate to
form ATP high- energy
compounds

ATP/ADP cycle

72
Cont.…
•ATP is ideal to transfer free energy from the
exergonic to the endergonic processes

•Hydrolysis of ATP produces more stable ADP and Pi

ATP + H2O ⇋ ADP ΔGo = –7.3

ATP + H2O ⇋ ADP

+ Pi kcal/mol
ΔGo = –7.7
kcal/mol
+ Pi

73
74
 Orthophosphate cleavage of ATP
•Removal of terminal γ-phosphate group from
ATP
 ATP + H2O → ADP + Pi, Δ G0= −7.3 kJ/mol
 Glutamate + NH3→ Glutamine ΔGo = 3.4 kJ/mol

•The excess ( 3.4 – 7.3= -3.9 kcal/mole) is

released as heat

75
 Pyrophosphate cleavage
•The β-and γ-phosphate groups are simultaneously
removed to form AMP & pyrophosphate (PPi)

76
 Other nucleoside triphosphates
•GTP, UTP, and CTP are energetically equivalent to
ATP

•Nucleoside diphosphate (NDP) kinases, can catalyze

the synthesis UTP,GTP, and CTP from their
diphosphates

•Serve in certain biosynthetic

pathways
 GTP →→ succinate
synthesis
 CTP →→ phospholipid 77
 Enzyme
• Enzymes speed up biochemical reaction by
lowering energy barriers

• The mechanism of enzyme action can be

viewed from two different perspectives.

 Energy changes

 Active site:

78
 Cont.….
Energy changes occurring during the reaction
• Every chemical reaction involves bond forming &
bond breaking, and requires energy is required to
begin the reaction

• Activation energy is energy difference between

reactants and high-energy intermediate (T*)

• The reactants must contain enough energy to

overcome the energy barrier of the transition state
(T*)

• An enzyme provides alternate reaction pathway 79

80
 Cont.….
Chemistry of the active site

• Active site is a complex molecular machine employing

a diversity of chemical mechanisms to facilitate the
conversion of substrate to product

• Two models have been proposed for substrate binding

 Lock-and-key model:- The enzyme active site

shape ⇒ rigid
has complementary 3-D shape to its substrate

 Induced-fit model : The active site of enzyme

substrate binds ⇒ flexible

undergoes a conformational change when the
81
Lock-and-key model Induced fit
model 82
 Cont.….
• Transition-state stabilization: Enzyme binds the
substrate and initiates its conversion to T*
 Increases the concentration of the reactive
intermediate

• Acid-base catalysis: amino acid residues in active site

provide or accept H+
 Histidine at the active site of chymotrypsin gains &
loses H+

• Nucleophilic catalysis a nucleophile is a chemical group

that is attracted to the positively charged nucleus 83
Reaction in the enzyme active
catalytic site 84
Cont.….
•Specificity
 Enzymes are highly specific, interacting with
one or a few substrates and catalyzing only one
type of chemical reaction
o Absolute specific ⇒ only single substrate

o Group specific ⇒ work with same functional group

o Linkage specific ⇒ catalyze specific combination of bond

o Stereochemically specific ⇒ only work with proper D or L- form

85
Cont.….
• Catalytic efficiency

 Most enzyme-catalyzed reactions are

highly efficient, proceeding from 103–108 x
faster than non-catalyzed reactions

 The number of molecules of substrate

converted to product per
enzyme per second, turnover rate (K cat )

o Typically, 100–104s-1

86
Cont.….
•Location within the cell
Many enzymes are localized in specific organelles
in the cell

 Compartmentalization isolates the reaction

substrate or product from other competing
reactions

Provides a favorable environment for the

reaction

 Organizes the thousands of enzymes into

purposeful pathways in cells 87
Factors affecting enzymes activities
• Enzymes can be isolated from cells & their
properties studied in a test tube (invitro)

• Different enzymes show different responses to

changes in local conditions:
o Substrate concentration
o Temperature
o pH
o Enzyme concentration
o Presence of inhibitor or activator

88
Cont.….
Substrate concentration
• The rate of an enzyme-catalyzed reaction
increases with substrate concentration increase
until a maximal
velocity (Vmax) is reached

89
Cont.….
Temperature
• Most enzymes function at body
temperature around 37 C0

• The reaction velocity increases wit

temperature until optimum
temperature,35-40

• Beyond optimum temperature rxn

velocity decreases due to enzyme
distortion

90
Cont.….
pH
• Every enzymes has its own range
of pH which function most
efficiently

 Most intracellular enzymes

best function at neutral
pH

 Some digestive enzymes

function at Acidic or basic pH

91
Inhibitor
• Inhibitor is any substance that can decrease the
rate of an enzyme-catalyzed reaction

Reversible
Competitive

Non-
Enzyme competitive
inhibitors
Uncompetiti
ve
Irreversibl
e
92
Cont.….
Reversible
• Where the activity can be restored to its original
function by the removal of the inhibitor
substance
non-covalent interactions
o hydrogen bonds
o ionic bonds
o hydrophobic interaction

93
Cont.….
Competitive Inhibitors
• Inhibitor binds active site &
competes with the substrate for
the site

94
Effect of a competitive inhibitor on the reaction velocity (vo) versus
95
substrate [S] plot
Cont.…. acetoacetyl-
acetyl-
Examples of competitive inhibition CoA CoA
• Statins
3-hydroxy-3-methylglutaryl-
Cholesterol lowering drugs CoA HMG-CoA
Statins reductase
 Competitive inhibitor of
HMG-CoA reductase Mevalona
te

Cholester
ol

96
Cont.….
Noncompetitive inhibitors
• The inhibitor substrate bind at
different sites on the enzyme

• The inhibitors can bind free

enzyme or the ES complex

98
A. Effect of a noncompetitive inhibitor on the reaction velocity (Vo) versus
99
substrate [S] plot. B. Lineweaver-Burk plot of noncompetitive inhibition of an
Cont.….
Examples of noncompetitive inhibitors
• Alanine inhibits private kinase none
competitively by bind other than active site
• Clinical importance of
noncompetitive inhibitors

Cyanide inhibits cytochrome

oxidase
Fluoride inhibits enolase

 Iodoacetate inhibits
glyceraldehyde-3- P
dehydrogenase

100
Cont.….
Uncompetitive inhibitors

Bind only to the [ES]

Inhibitor does not resemble to
the S
Vmax & Km decrease

101
Cont.….
Irreversible inhibitors
• Covalently bound to the essential groups of
enzymes
• Cause a partial or complete loss of the
enzymatic activity
• Loss of activity cannot be recovered within the
stipulated time of interest

102
Cofactors &
Coenzymes
• Apoenzyme
 The protein part of an
enzyme

• Coenzyme and cofactor

Nonprotein moiety

⇒ FAD, NAD+
•Coenzyme (organic molecule)

⇒ Zn2+,Fe2+
•cofactor (metal ion)

103
Cont.….
• Holoenzyme
Active enzyme with its nonprotein
component

104
Cont.….
Cofactors
• Associate directly with the enzyme or in the form of a
cofactor-
substrate complex

• Metal-activated enzymes: metal ion loosely bind

• Metalloenzymes: metal ion tightly bind

• Function
Mediate oxidation reduction reaction

Properly orient substrates for reaction

Electrostatically stabilize negative charges

105
Cont.….

Cofactor Function Enzyme

Mg2+ Substrate binding Kinases
Ca2+ Substrate Hydrolase
activation
K+ Stabilization Pyruvate
kinase
Fe2+ Electron transport Oxidase
Zn2+ Substrate binding DNA
polymerase
Cu+ Electron transport Oxidase
Cont.….
• Mg2+ in
hexokinase

Mg2+ interacts with negatively charged phosphate oxygen atom, providing charge
composition &
promoting favorable conformation of at active site
107
Cont.….
• Copper and Iron in cytochrome
oxidase

Cytochrome oxidase is a metallo-protein ,

containing copper and iron atoms,

108
Cont.….
• Coenzymes
 Are mixed bag of organic compounds; vitamins or
metabolically related to vitamins

Most important coenzymes are especially B vitamins

 Act to transfer chemical groups between enzymes or

from enzyme to substrate or product

 Transport variety of chemicals groups such as hydride,

acetyl, formyl,
methenyl, methyl…

e.g. NAD+,FAD
109
Cont.….
• Two type of coenzymes
Cosubstrates are altered during the
reaction and regenerated by another
enzyme

e.g. NAD+

Prosthetic groups remain bound to the enzyme

during the reaction, and may be covalently
bound to enzyme

e.g. FAD, biotin

110
111
Cont.….
•The coenzyme NAD+ accepting a
hydride ion from lactate

•NAD-dependent dehydrogenases
catalyze the transfer of a
hydride ion (H-) from a carbon to
NAD+ in oxidation reactions

112
 Regulation of enzyme activity
•Enzyme activities can be regulated
(activated or inhibited

• Rate of product formation responds to the

needs of the cells
• Three mechanisms are proposed
Allosteric regulation e.g. feed back inhibition
Covalent modification e.g. phosporylation
Induction & repression of enzyme synthesis

11
3
Cont.….
Allosteric regulation
• Allosteric enzymes are regulated by a
molecules called effectors/modifiers

• Bind noncovalently at a site other than the

active site

• Effectors- are either positive (accelerate) or

negative (slow down)
the reaction

• Allosteric effector can alter the affinity of enzyme for

its substrate, or modify maximal catalytic activity
of enzyme, or both
11
4
Effects of negative or positive effectors on an allosteric enzyme

11
5
Cont.….
Regulation of enzymes by covalent modification
• Many enzymes may be regulated by covalent
modification, most frequently by the addition or
removal of phosphate groups

• For example phosphorylation of:

glycogen phosphorylase
(active)
11
glycogen synthase 6
Cont.….
Induction & repression of enzyme synthesis
• Cells can also regulate the amount of enzyme present
by altering the rate of enzyme synthesis

• Increase (induction) or decrease (repression) of enzyme

synthesis leads to an alteration in the total
population of active sites

• For example elevated levels of insulin cause an

increase in the
synthesis of key enzymes involved in glucose
metabolism

11
7
 Clinical applications of enzymes
• Therapeutic uses:
 More than half of drugs prescribed for
different disease targeted enzyme

 Such as
 Allopurinol (Rx of gout) targeted xanthine
oxidase
 Aspirin (anti-inflammatory) targeted
cyclooxygenase
 Methotrexate (anticancer) targeted
dihydrofolate reductase
11
8
Cont.….
• Enzymes in clinical diagnosis
 Plasma enzymes can be classified into two major
groups.

‐ Enzymes actively secreted into the blood by

certain cell types. e.g. liver secretes blood
clotting zymogens

‐ Enzyme species released from cells during

normal cell turnover
* These enzymes almost always function
intracellularly & have no physiologic use in the
plasma 11
9
Release of enzymes from normal and diseased or traumatized
cells

120
Cont.….
• Alteration of plasma enzyme levels in disease states
 Diseases that cause tissue damage result in an
increased release of intracellular enzymes into
the plasma

 The activities of these enzymes are routinely

determined for diagnosis diseases (heart, liver,
skeletal muscle, & other tissues)

 The level of specific enzyme activity in the

plasma frequently correlates with the extent
of tissue damage
121
Cont.….
• Plasma enzymes as diagnostic tools
 Some enzymes show relatively high activity in
only one or a few tissues

 The presence of increased levels of these

enzymes in plasma thus reflects damage to the
corresponding tissue

 Alanine aminotransferase (ALT) is abundant in

the liver

122
Cont.….
• Isoenzymes & diseases of the heart enzymes
 Isoenzymes are enzymes that catalyze the same
reaction
 However, they do not necessarily have the same
physical properties due to differences in amino
acid sequence
 Isoenzymes may contain different numbers of
charged aas & may be separated from each
other by electrophoresis
 Different organs frequently contain
characteristic proportions of different 123
Cont.….
 Different organs frequently contain
characteristic proportions of different
isoenzymes

 The pattern of isoenzymes found in the plasma

may serve as a means of identifying the site of
tissue damage

 Creatine kinase (CK) occurs as three isoenzymes

 Lactate dehydrogenase occurs as 5 isoenzymes

124
Isoenzymes of
CK
Isozyme Composition Present in

CK-1 BB Brain
CK-2 MB Myocardium
CK-3 MM Skeletal
muscle

125
Isoenzymes of LDH
Isoenzyme Composition Present in

LDH-1 HHHH Myocardium

LDH-2 HHHM Myocardium, RBC
LDH-3 HHMM Kidney, skeletal
muscle
LDH-4 HMMM Kidney, skeletal
muscle
LDH-5 MMMM Skeletal muscle

126
Cont.….

 Myocardial muscle is the only tissue that contains

more than 5% of the total CK activity as the CK2
(MB) isoenzyme

 Appearance of this hybrid isoenzyme in plasma is

virtually specific for infarction of the
myocardium

 Following an acute MI, CK2 appears 4–8 hours,

reaches a peak of activity at approximately 24
hrs, and returns to baseline after 48–72 hours 127
128