AMYA

POLYTECHNIC COLLEGE
Bonum Est Sui Diffusivum

Biochemistry for MLS

Course No.: MT 201
Course Title: Biochemistry for Medical Laboratory Science
Unit: 5 Units (3 Unit Lecture; 2 Units Laboratory)
Pre-/Co-Requisite: MT 105 and MT 107
Year Level: 2nd Year, 1st Semester

MEDICAL LABORATORY SCIENCE PROGRAM

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 Table of Contents

page

Module 1 Overview of Biochemistry 4

Module 2 Biochemical Energy Production 6
Module 3 Carbohydrates 9
Module 4 Lipids 16
Module 5 Proteins 20
Module 6 Enzymes 28
Module 7 Nucleic Acid 30

- JB SANCHEZ

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 AMYA
POLYTECHNIC COLLEGE MEDICAL LABORATORY
Bonum Est Sui Diffusivum SCIENCE PROGRAM

COURSE MODULE
Biochemistry for Medical Laboratory Science
Course No.: MT 201
Course Title: Biochemistry for Medical Laboratory Science
Unit: 5 Units (3 Unit Lecture; 2 Units Laboratory)
Pre-/Co-Requisite: MT 105 and MT 107
Year Level: 2nd Year, 1st Semester

Course Description:
This course deals with the structure, classifications and functions of the different
biochemical substances in the body. This includes the chemistry of carbohydrates , lipids,
proteins ,enzymes , nucleic acids and its metabolism.

Course Objectives:
At the end of the course, the learners are expected to:

Cognitive
1. Define common terms used in biochemistry
2. Describe the characteristics of simple and more complex organic compounds
commonly involved in biological reactions
Affective

1. Appreiate the relevance of the biochemistry concepts to everyday lif existence by

recognizing the events in which the principles are clearly manifested

Psychomotor

1. Detect disparities in the standard technique, if there is any, and propose modification
through research.
2. Assume a stance in truth of every situation in phlebotomy regardless of personal interests

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Learning Module No. 1

Topic: Overview of Biochemistry

Materials: Laptop and Internet Connection
Assessment: MCQ Post-Test via Infinit

Learning Outcomes:
At the end of the session, the students should be able to:
1. Explain the basic information and relevance of biochemistry.
2. Define common terms used in biochemistry

Biochemistry – Meaning and Scope

1. Meaning of Biochemistry
Biology in its broadest sense may be regarded as the study of the biosphere,
which consists of organisms of all types and the environment in which they live.
Biochemistry is the chemistry of life and its environment. Biochemistry is the science
concerned with studying the various molecules that occur in living organisms and their
environment, with their chemical reactions and processes they undergo. In other words,
biochemistry deals with the structure and function of the body at the molecular level.
The aim of biochemistry is to describe and explain, in molecular terms, all
chemical processes of living organisms and their interactions with their environments
both in health and disease conditions.
2. Scope of Biochemistry
The scope of biochemistry is as wide as life itself. Wherever there is life,
chemical processes are occurring. Biochemists study the chemical processes that occur
in microorganisms, plants, insects, fish, birds, mammals and human beings. Students in
the biomedical sciences will be particularly interested in the biochemistry of the two latter
groups.
Biochemistry is concerned with the entire spectrum of life forms, from relatively
simple viruses and bacteria to complex human being. Even evolution of life, which
started with chemical evolution, works at the molecular level (at the level of DNA). Living
organisms are enormously diverse in appearance and function. Birds, beasts, trees,
grasses and microscopic organisms differ greatly. Yet, biochemical research has
revealed that all organisms are remarkably alike at the cellular and chemical levels.
Biochemistry is the language of biology. The tools for research in all the branches of
medical science are mainly biochemical in nature. Because life depends on biochemical
reactions, biochemistry has become the basic language of all biologic sciences. In fact,
the old barriers among the life sciences are breaking down and biochemistry is
increasingly becoming their common language.
Genetic Engineering (DNA technology/recombinant DNA techniques), a branch
of biochemistry, is the most advanced and sophisticated tool of biotechnology. Genetic
engineering has applications in the fields of medicine, agriculture, animal farming,
ecology, paleontology, etc.
Advances in biochemistry are enabling researchers to tackle some of the most
exciting questions in biology. How life evolved on earth about 4 billion years ago? How a
single molecule of DNA replicates to generate two identical copies of itself? How does a
cell divide? How does a fertilized egg give rise to cells as different as those in muscle,
brain and liver? How do the senses work? How does the immune system distinguish
between self and non-self? What are the molecular mechanisms of memory and

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 behavior? Which molecules are involved in processes like sleep, aggression and various
moods and emotions?

3. Medical Biochemistry
Medical biochemistry deals with the chemical aspects of both health and
diseases of human beings. Normal biochemical processes are the basis of health.
Conversely, every disease has a biochemical basis. That is, all diseases are
manifestations of abnormalities of molecules, chemical reactions, or processes. A
knowledge of biochemistry is essential to the understanding of all branches of medical
science, especially, physiology, immunology, pharmacology, pharmacy, toxicology,
pathology, general medicine, psychiatry, etc.
In pharmaceutical industry, for example, biochemistry is useful in the rational
design of new drugs – e.g. specific inhibitors of enzymes required for the replication of
viruses such as human immunodeficiency viruses (HIV)
Techniques of genetic engineering/DNA technology/recombinant DNA
techniques, the most advanced branch of biochemistry, have many medical applications:
Basic research for understanding structure and functions of DNA and proteins
Diagnosis of genetic and microbial diseases (e.g. aids)
Forensic applications (e.g. solving disputes of parenthood by DNA fingerprinting)
Production of proteins (in pharmaceutical industry) for:
a) Replacement therapy (e.g. insulin)
b) Disease prevention (e.g. vaccines)
c) Diagnostic tests (e.g. monoclonal antibodies).
d) Treatment of genetic diseases (e.g. gene therapy)
Most important use of medical biochemistry, however, is biochemical tests done
in the clinical laboratory.
The uses of biochemical tests are in – 1) diagnosis, 2) monitoring and 3)
screening of diseases
1. Diagnosis – confirming or ruling out clinical diagnosis and assessment of
severity/extent of diseases and prognosis
2. Monitoring – following the progress of disease and its response to treatment
3. Screening – to detect early disease or risk factors to develop disease
Conclusions
Broadly, uses of biochemistry for medical science are as follows:
1. To understand and reveal the fundamental causes and mechanisms of
disease processes.
2. To assist the diagnosis, monitoring and screening of specific diseases by the
judicious use of various biochemical laboratory tests.
3. To suggest rational strategy for the treatment and prevention of diseases.

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Learning Module No. 2

Topic: Biochemical Energy Production

Materials: Laptop and Internet Connection
Assessment: MCQ Post-Test via Infinit

Learning Outcomes:
At the end of the session, the students should be able to:
1. Discuss the cell structure and its importance in the metabolism
2. Explain the energy production in the body and its pathway

METABOLISM
- sum total of all the biochemical reactions that take place in a living organism
- an average human adult whose weight remains the same for 40 years processes about
6 tons of solid food and 10,000 gallons of water
Two Subtypes of Metabolic Reaction:
A. catabolism
- all metabolic reactions in which large biochemical molecules are broken down
to smaller ones
- usually release energy
- involved in the oxidation of glucose
B. anabolism
- all metabolic reaction in which small biochemical molecules are joined together
to form larger ones
- usually require energy
- ex. Synthesis of proteins from amino acids

METABOLIC PATHWAY
- a series of consecutive biochemical reactions used to convert a starting material into
an end product
- major metabolic pathways for all life forms are similar which enables scientists to study
metabolic reactions in simpler life forms and use the results to help understand the
corresponding metabolic reactions in more complex organisms like humans
- linear: a series of reactions generates a final product

IMPORTANT INTERMEDIATE COMPOUNDS IN METABOLIC PATHWAYS

- All have nucleotides

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AMP
- one of the nucleotides present in RNA molecules
- ATP and ADP differ structurally from AMP only in the number of phosphate groups
present

ATP
- a phosphoester bond joins the first phosphoryl group to the pentose sugar ribose
- the other two phosphoryl groups are joined to one another by phosphoanhydride bonds

PHOSPHORYL GROUP
- functional group derived from a phosphate ion that is part of another molecule
- ATP: 3; ADP: 2; AMP: 1 phosphoanhydride bond
- chemical bond formed when two phosphate groups react with each other and a water
molecule is produces
- when two phosphate groups react with one another, a water molecule is produced hence
“anhydride”

- ATP and ADP molecules readily undergo hydrolysis reactions in which phosphate groups (Pi,
inorganic phosphate) are released
- in metabolic pathways in which they are involved, the adenosine phosphates continually
change back and forth among the various forms
- these hydrolyses are energy-producing reactions that are used to drive cellular processes that
require energy input
- phosphoanhydride bonds in ATP and ADP are very reactive bonds that are require less energy
than normal to break

HIGH ENERGY PHOSPHATE DEMANDS

1. high-energy compound
- compound that has a greater free energy of hydrolysis than that of a typical compound
- contain one or more very reactive bonds, strained bonds
- the balance between the energy needed to break bonds in the reactants and that
released by bond formation in the products > typical amount of free energy released
during hydrolysis
2. free energy
- amount of energy released by a chemical reaction that is actually available for further
use (unlike heat)

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In chemical reaction, the energy balance between the bond breaking among reactants (energy
input) and new bond formation among products (energy release) determines whether there is a
net loss or a net gain of energy.

3. H3PO4
- weak triprotic inorganic acid
- parent molecule for phosphate groups
- exists in aqueous solution in several forms
- dominant form

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Learning Module No. 3

Topic: Carbohydrates
Materials: Laptop and Internet Connection
Assessment: MCQ Post-Test via Infinit

Learning Outcomes:
At the end of the session, the students should be able to:
1. Recognize and understand how carbohydrates produced in the body
2. Understand the complex function of the carbohydrates
3. Define and classify carbohydrates based on composition and structure
4. List the appropriate methods in quantifying carbohydrates

INTRODUCTION

Carbohydrates (from 'hydrates of carbon') or saccharides (Greek σάκχαρον meaning "sugar")

are simple organic compounds that are aldehydes or ketones with many hydroxyl groups added,
usually one on each carbon atom that is not part of the aldehyde or ketone functional group.

All carbohydrates can be classified as either monosaccharides, oligosaccharides or

polysaccharides. Anywhere from two to ten monosaccharide units, linked by glycosidic bonds,
make up an oligosaccharide. Polysaccharides are much larger, containing hundreds of
monosaccharide units. The presence of the hydroxyl groups allows carbohydrates to interact with
the aqueous environment and to participate in hydrogen bonding, both within and between chains.
Derivatives of the carbohydrates can contain nitrogens, phosphates and sulfur compounds.
Carbohydrates also can combine with lipid to form glycolipids or with protein to form glycoproteins.

Carbohydrate Nomenclature

The predominant carbohydrates encountered in the body are structurally related to the
aldotriose glyceraldehyde and to the ketotriose dihydroxyacetone. All carbohydrates contain at
least one asymmetrical (chiral) carbon and are, therefore, optically active. In addition,
carbohydrates can exist in either of two conformations, as determined by the orientation of the
hydroxyl group about the asymmetric carbon farthest from the carbonyl. With a few exceptions,
those carbohydrates that are of physiological significance exist in the D-conformation. The mirror-
image conformations, called enantiomers, are in the L-conformation.

Classification of Carbohydrates

Monosaccharides
The monosaccharides commonly found in humans are classified according to the number of
carbons they contain in their backbone structures. The major monosaccharides contain four to

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six carbon atoms. Monosaccharides are the simplest carbohydrates in that they cannot be
hydrolyzed to smaller carbohydrates. The general chemical formula of an unmodified
monosaccharide is (C•H2O)n, where n is any number of three or greater.

Different Type Of Monosaccharides

CATEGORY
CARBONS RELEVANT EXAMPLES
NAME

3 Triose Glyceraldehyde, Dihydroxyacetone

4 Tetrose Erythrose

5 Pentose Ribose, Ribulose, Xylulose

6 Hexose Glucose, Galactose, Mannose, Fructose

7 Heptose Sedoheptulose

9 Nonose Neuraminic acid also called sialic acid

Glucose

(Glc), a monosaccharide (or simple sugar), is an important carbohydrate in biology. The living cell
uses it as a source of energy and metabolic intermediate. Glucose is one of the main products of
photosynthesis and starts cellular respiration in both prokaryotes and eukaryotes. The name
comes from the Greek word glykys (γλυκύς), which means "sweet", plus the suffix "-ose" which
denotes a carbohydrate
Glucose is commonly available in the form of a white substance or as a solid crystal. It can also
be commonly found as an aqueous solution

Prepration

1. From Sucrose
If sucrose is boiled with dilute HCL or H2SO4 in alcoholic solution glucose and fructose are
obtained in equal amount
C12H22O11 + H2O C6H12O6 + C6H12O6
glucose fructose

2. From Starch
Glucose is obtained by hydrolysis of starch by boiling it with dilute H2SO4 at 393 K under
pressure.
H+
{ C6H10O5 }n + nH2O C6H12O6
393 k – 2-3 atm
Glucose

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 Structure
Glucose (C6H12O6) contains six carbon atoms one of which is part of an aldehyde group and is
therefore referred to as an aldohexose. The glucose molecule can exist in an open-chain (acyclic)
and ring (cyclic) form (in equilibrium), the latter being the result of an covalent bond between the
aldehyde C atom and the C-5 hydroxyl group to form a six-membered cyclic hemiacetal. In water
solution both forms are in equilibrium, and at pH 7 the cyclic form is predominant. As the ring
contains five carbon atoms and one oxygen atom, which resembles the structure of pyran, the
cyclic form of glucose is also referred to as glucopyranose. In this ring, each carbon is linked to a
hydroxyl side group with the exception of the fifth atom, which links to a sixth carbon atom outside
the ring, forming a CH2OH group.

Glucose Structure

Cyclic Structure of Glucose

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Fructose
Fructose (or levulose) is a simple reducing sugar (monosaccharide) found in many foods and is
one of the three most important blood sugars along with glucose and galactose. Honey, tree fruits,
berries, melons, and some root vegetables, such as beets, sweet potatoes, parsnips, and onions,
contain fructose, usually in combination with sucrose and glucose. Fructose is also derived from
the digestion of sucrose, a disaccharide consisting of glucose and fructose that is broken down
by glycoside hydrolase enzymes during digestion. Fructose is the sweetest naturally occurring
sugar, estimated to be twice as sweet as sucrose.

Structure of Fructose

Cyclic Structure of Fructose

Disaccharides

Covalent bonds between the anomeric hydroxyl of a cyclic sugar and the hydroxyl of a second
sugar (or another alcohol containing compound) are termed glycosidic bonds, and the resultant
molecules are glycosides. The linkage of two monosaccharides to form disaccharides involves a
glycosidic bond. Several physiogically important disaccharides are sucrose, lactose and maltose.

• Sucrose: prevalent in sugar cane and sugar beets, is composed of glucose and fructose
through an -(1,2) -glycosidic bond.

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 Sucrose

• Maltose: the major degradation product of starch, is composed of 2 glucose monomers

in an -(1,4) glycosidic bond.

• Lactose: is found exclusively in the milk of mammals and consists of galactose and
glucose in a -(1,4) glycosidic bond.

Polysaccharides

Most of the carbohydrates found in nature occur in the form of high molecular weight polymers
called polysaccharides. The monomeric building blocks used to generate polysaccharides can be
varied; in all cases, however, the predominant monosaccharide found in polysaccharides is D-
glucose. When polysaccharides are composed of a single monosaccharide building block, they
are termed homopolysaccharides. Polysaccharides composed of more than one type of
monosaccharide are termed heteropolysaccharides.

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Starch
Starch is the major form of stored carbohydrate in plant cells. Its structure is identical to glycogen,
except for a much lower degree of branching (about every 20-30 residues). Unbranched starch is
called amylose; branched starch is called amylopectin.

Glycogen
Glycogen is the major form of stored carbohydrate in animals. This crucial molecule is a
homopolymer of glucose in -(1,4) linkage; it is also highly branched, with -(1,6) branch linkages
occurring every 8-10 residues. Glycogen is a very compact structure that results from the coiling
of the polymer chains. This compactness allows large amounts of carbon energy to be stored in
a small volume, with little effect on cellular osmolarity.

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Cellulose
Cellulose is an organic compound with the formula (C6H10O5)n. It is a structural polysaccharide
derived from beta-glucoseCellulose is the primary structural component of green plants. The
primary cell wall of green plants is made of cellulose; acetic acid bacteria are also known to
synthesize cellulose, as well as many forms of algae, and the oomycetes. Cellulose was
discovered and isolated in the mid-nineteenth century by the French chemist Anselme Payenand,
as of the year 2006, the estimated annual production is 1.5x109 tonnes. Some animals,
particularly ruminants and termites, can digest cellulose with the help of symbiotic micro-
organisms (see methanogen). Cellulose is not digestible by humans and is often referred to as
'dietary fiber' or 'roughage', acting as a hydrophilic bulking agent for feces.

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Learning Module No. 4

Topic: Lipids
Materials: Laptop and Internet Connection
Assessment: MCQ Post-Test via Infinit

Learning Outcomes:
At the end of the session, the students should be able to:
1. Identify the general properties of lipids and fatty acids
2. Classify lipids
3. Discuss the role of triacylglycerol, sphingogycolipids and cholesterol in the body
4. Describe the digestion, absorption of carbohydrates in the body
5. List the appropriate methods in quantifying Lipds

Lipids in general
Lipids are insoluble in water but soluble in organic solvents, like chloroform
Lipids are made up of carbon, hydrogen and oxygen
Lipids have no generalized formula and much less oxygen compared to hydrogen
Lipids have a high proportion of non-polar carbon-hydrogen bonds, making them
hydrophobic
Fat is a lipid in the solid state at 20oC
Oil is a lipid in the liquid state at 20oC
Simple lipids

Simple lipids are esters( )of fatty acids with various alcohol
Fats are esters of fatty acids with glycerol.
• Monoglyceride = 1 fatty acid molecule + 1 glycerol molecule
• Diglyceride = 2 fatty acid molecules + 1 glycerol molecule
• Triglyceride = 3 fatty acid molecules + 1 glycerol molecule

Waxes are esters of fatty acids with complex alcohols

Compound lipids
Compound lipids = lipid + non-lipid components
• Phospholipids = lipid + phosphate
• Glycolipids = lipid + carbohydrate
• Steroids consists of a complex ring of carbon atoms

Structure of lipids
Lipids are composed of glycerol and fatty acids
• Glycerol is an alcohol with three carbons, each bearing a hydroxyl group (-OH)

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 • Fatty acids consist of a hydrocarbon chain and a carboxyl group, with the general

formula RCOOH, where R is a hydrocarbon chain.

o A hydrocarbon chain consists of a chain of carbon atoms bonded to hydrogen
atoms
§ They vary in length, but a usually even numbered chains of 14 to 22
carbons in length
o These hydrocarbon chains may be saturated or unsaturated, causing the fatty
acid to be saturated or unsaturated
§ A saturated hydrocarbon chain does not contain any C=C and contains
the maximum number of hydrogen atoms possible
v This gives rise to saturated fatty acids and saturated fats
v Fats that are more saturated (contain more saturated fatty acids)
have a higher melting and boiling point, causing them to be
more solid
v All animal fats are saturated
§ An unsaturated hydrocarbon chain contains C=C, giving rise to kinks in

the chain.
v If more than one C=C is present, it is polyunsaturated
v This gives rise to unsaturated fatty acids and unsaturated fats
v Fats that are unsaturated have a lower melting and boiling
point, as the kinks prevents them from packing tightly with each
other, hence causing them to be usually liquid at room
temperature
v Most plant fats are unsaturated

Synthesis of lipids
Lipids are formed from the condensation reaction between glycerol and fatty acid molecules.
In the condensation reaction, each hydroxyl group (-OH) in the glycerol molecule reacts with
the carboxyl group (-COOH) of the fatty acid, producing one water molecule and an ester

bond ( ).
Condensation reaction = when two compounds are joined by the elimination of water
molecule(s)
Phospholipids
Phospholipids form the core of all biological molecules, and are made up of three kinds of
subunits:
• Glycerol
o Forms the backbone of the phospholipid molecule

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 • Fatty acids
o Two fatty acids are attached
• Phosphate group

o A charged phosphate group ( ) is attached to one end of the glycerol

molecule, usually with a charged organic molecule attached to it

The simplest phospholipid is phosphatidic acid, consisting of a glycerol molecule, 2 fatty acid
molecules and one phosphoric acid.
Phospholipids are amphipathic, as they have a charged/ polar hydrophilic phosphate head
with a non-charged/ non-polar hydrophobic fatty acids tails
Steroids
There are many important steroids like sex hormones and cholesterol
Cholesterol is highly hydrophobic (not amphipathic), as it only has a single hydroxyl group
(negligible compared to the rest of the carbon rings). It is also the most abundant steroid in
animals and is largely absent in plants
Functions of triglycerides
Function of triglycerides Structure Structure-function
relationship
As an efficient storage 1 triglyceride molecule Triglycerides can be stored in
molecule of energy consists of three fatty acids large amounts without
molecules attached to a having any effect on the
glycerol molecule by ester water potential of cells
bonds
They also cannot diffuse out
The triglyceride is large and of the cells while insoluble
uncharged, causing it to be
insoluble in water Hence making them the ideal
storage molecule
As compared to carbohydrates of similar mass, the ratio of
energy storing C-H bonds in triglycerides is more than twice
that of carbohydrates
Hence, as compared to the similar mass of carbohydrates, fat
yields more chemical energy then carbohydrates

Also, for the same amount of energy to be produced, less

than half the mass of triglycerides are needed. For animals,
this means less mass, allowing for quicker locomotion. For
plants, seeds can be kept small and light, making dispersal
easier
As an important source of For the same mass of carbohydrates, triglycerides contain
metabolic water twice the number of hydrogen atoms, which forms the
metabolic water when oxidized in respiration

Hence, triglycerides produce twice as much metabolic water

as carbohydrates when oxidized in respiration

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 This is important for dessert animals (like camels) who
survive off metabolic water
Provide buoyancy Triglycerides are less dense Allows aquatic mammals to
than water stay buoyant in water
As long-term energy store Triglycerides are only oxidized after carbohydrates are
depleted

This is important for hibernating animals

As thermal insulator Triglycerides prevent excessive heat loss

This is important for mammals living in cold climates and

aquatic mammals
Protecting delicate organs Triglycerides surround the organs and serves to absorb
shock

Functions of lipids in cell membrane

Cholesterol, glycolipids and phospholipids are important components of the cell membrane
• Cholesterol
o Maintains fluidity of cell membranes, as it disturbs the close packing of
phospholipids (like the kinks in the fatty acid), thus keeping them more fluid
• Glycolipids
o The carbohydrate attached is used for cell recognition and cell adhesion
• Phospholipids
o Major constituent of cell membrane, as they form the bilipid layer of the cell
membrane, forming a hydrophobic boundary between the aqueous interior
and exterior of the cell
o This is due to the amphipathic nature of the phospholipids, arising from the
hydrophilic, charged phosphate head and the hydrophobic hydrocarbon tails
o This result in them forming a bilayer in an aqueous environment, with the
phosphate heads facing outwards, in contact with the aqueous environment on
either side.
o Whereas the hydrophobic tails are facing inwards and are buried between the
hydrophobic heads
o Therefore, the hydrophobic region of the bilayer forms the boundary between the
aqueous interior and exterior of the cell
o The hydrophilic region allows the hydrophobic boundary to exist in an aqueous
environment

Miscellaneous functions
Cholesterol is used in the synthesis of steroid hormones and vitamin D3
Lipids are also components of myelin sheath nerve cells, where they act as electrical insulators,
to allow rapid transmission of impulses along myelinated nerves

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Learning Module No. 5

Topic: Proteins
Materials: Laptop and Internet Connection
Assessment: MCQ Post-Test via Infinit

Learning Outcomes:
At the end of the session, the students should be able to:
1. Recall the structure and characteristics of proteins
2. Explain how the body metabolize, digest and reabsorbs proteins in the body.
3. List the appropriate methods in quantifying proteins

PROTEINS
- Performs many vital functions in the body such as:
- Structural support; enzymes; movement; transport; recognition and receptor molecules;
regulation of proteins and DNA; hormones; antibodies; toxins and venoms
- The monomer of proteins is called AMINO ACID.

AMINO ACID
- Organic compounds containing an amine and carboxylic group and a carbon chain
specific to an amino acid.

Amino acid general

structure

- Amino acids bond covalently to form peptides and polypeptides

- Peptide bonds happen between the terminal carboxyl of one amino acid and terminal
amino group of the next amino acid.
- A dehydration synthesis reaction occurs and forms the peptide bond.
- Disulfide bond happens between two terminal sulfhydrils of different amino acids

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TYPES OF PROTEINS
1. Primary structure is the unique sequence of amino acids forming a polypeptide
- Primary structure of a protein is the precise sequence in which amino acids are linked
- Changing even a single amino acid alters secondary, tertiary, and quaternary structures
– which can alter or destroy the biological function of a protein
- Example: Substitution of a single amino acid in hemoglobin produces an altered form
responsible for sickle-cell disease

2. Secondary structure is produced by the twists and turns of the amino acid chain

- The amino acid chain (primary structure) is folded into arrangements that form the
protein’s secondary structure
- Amino acid side groups extend outward from the twisted backbone

ARRANGEMENT OF SECONDARY STRUCTRES:

• The alpha (α) helix is twisted into a regular right-hand spiral
• The beta (β) strand/pleated sheet zigzags in a flat plane, forming a sheet

alpha (α) helix beta (β) strand/

pleated sheet

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3. Tertiary structure is the folding of the amino acid chain, with its secondary structures,
into the overall 3Dshape of a protein
- Tertiary structure gives a protein its overall three-dimensional shape, or conformation
- The positions of secondary structures, disulfide linkages, and hydrogen bonds play
major roles in folding each protein into its tertiary structure
- Attractions between positively and negatively charged side groups and polar or nonpolar
associations also contribute to tertiary structure
- Tertiary structure determines a protein’s function
- The distribution and 3-D arrangement of side groups, in combination with their chemical
properties, determine the overall chemical activity of the protein
- Tertiary structure also determines the solubility of a protein, depending on the
arrangement of polar (hydrophilic) and nonpolar (hydrophobic) segments Tertiary
structure of most proteins is flexible, allowing them to undergo limited conformational
changes
- Conformational changes are important to the function of enzymes, and to proteins
involved in cellular movements or transport of substances across cell membranes

- DENATURATION - Unfolding a protein from its active conformation so that it loses its
structure and function (caused by chemicals, changes in pH, or high temperatures)
- For some proteins, denaturation is permanent – for others, denaturation is reversible
(renaturation)

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4. Quaternary structure – when present, is formed from more than one polypeptide chain
- Some complex proteins, such as hemoglobin and antibody molecules, have quaternary
structure the presence and arrangement of two or more polypeptide chains
- Hydrogen bonds, polar and nonpolar attractions, and disulfide linkages hold the multiple
polypeptide chains together
- In many proteins, folding of the amino acid chain (or chains) produces large subdivisions
called domains
- In proteins with multiple functions, individual functions are often located in different
domains
- Domains with similar functions are found in different proteins
- 3-D arrangement of amino acid chains within and between domains produces highly
specialized regions called motifs

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PROTEINS

Consist of: C, H, O, N and sometimes S

Basic structural unit: amino acid
Proteins differ from Carbs: ALWAYS CONTAIN NITROGEN

AMINO ACIDS
• >100 naturally occurring, only 20 used in biosynthesis
• Consists of:
o a-carbon atom (which is bonded to)
o Hydrogen atom
o Amino group (-NH2)
o Carboxyl group (-COOH)
o Variable R group (aka side chain)
§ Different amino acids have different R groups

Classification of amino acids

(A) Based on human nutrition
a. Essential amino acids
i. Cannot be synthesized from simple substances by human
ii. Must be obtained from diet
b. Non-essential amino acids
i. Can be synthesized from simpler substances
(B) Properties of R group
a. Neutral amino acids
i. Have non-polar (hydrophobic) or polar (hydrophilic) side chains
ii. Sum of +ve and –ve charges are equal
b. Electrically-charged amino acids
i. Negatively-charged: acidic amino acids (hydrophilic)
ii. Positively charged: basic amino acids (hydrophilic)

Properties of amino acids

(I) Exists as zwitterions (aka dipolar ion)
a. Not ALL the time though!
b. In solution, ionized a.a. carry both +ve and –ve charges
i. Protonated: (-NH2) receives H+ à (-NH3+)
ii. Deprotonated: (-COOH) dissociates H+ à (-COO-)
(II) Amino acids as buffers
a. Amphoteric: can act as both acids and bases à good buffer quality
b. Buffer: prevents changes in pH when small amt of acid/alkali is added à by
accepting or donating H+
i. When acid (H+) is added to the solution, COO- of zwitterion accepts a H+
to neutralize the H+, and becomes COOH
Thus preventing a change in pH of the solution, a.a. becomes positively
charged
(III) Based on R groups of amino acids
a. Physical & chemical properties determine uniqueness of a.a. à
Charge/size/shape/reactivity

POLYPEPTIDES

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 • Monomers: amino acids
o Dipeptide (2 residues)
o Tripeptide (3 residues)
o Oligopeptide (3-10 residues)
o Polypeptides (>10 residues)
• Formed through condensation/dehydration
o Links carboxyl group to amino group, removes water molecule
o Peptide bond!
• Backbone: regularly repeating part, main chain
• Variable part: distinctive vatiable R groups
• Amino-terminal (N-terminal) à start of polypeptide chain
Carboxyl-terminal (C-terminal) à end of polypeptide chain
• Folds into specific 3-D shape

STRUCTURE OF PROTEINS
Primary Structure
Refers to sequence and number of amino acids in a polypeptide chain.
• Bond: peptide bonds between successive a.a.
• Each polypeptide chain has unique sequence/number/type of a.a.
o Sequence determines type and location of cross linkages à pattern of folding à
unique 3-D conformation
o NOT random! à linking of a.a. determined by nucleotide sequences in genes
• Important in determining the function of the protein
o One amino acid alteration in the primary structure can alter the entire structure of
the protein
o Sequence of nucleotides in the gene + sequence of amino acid residues =
important in determining shape and function
o E.g. sickle cell anaemia à glutamic acid (6th a.a.) replaced with valine à results
in abnormal b-globin polypeptide à alters entire structure of the haemoglobin à
altered property (haemo molecules aggregate together at low oxygen
concentrations à causes red blood cells to change shape
• Primary structure remains unaffected during denaturation

Secondary Structure
Refers to spatial arrangement formed by regular coiling and pleating of a single polypeptie chain
• Bond: H bonds at regular intervals (between CO and NH grps of backbone)
• a-helix: single polypeptide chain coiled into an extended spiral spring
o CO groups of one turn linked to NH grps of the next turn (ALL CO & NH are
involved à stability) at every 4th peptide bond
o 3.6 amino acid residues in every turn
o E.g. keratin (structural protein of hair), wool, nails
• b-pleated sheet: when 2 or more regions/segments of a single polypeptide chain lying
side-by-side are linked by H-bonds
o NH & CO grps of one chain will form bonds with NH & CO grps from adjacent
chain (ALL involved à structure is stable and rigid)
o Runs parallel (same) or anti-parallel (opposite) à forms a flat sheet which
becomes folded
o R-groups are not involved in bonding à project above or below the plane
o E.g. Fibroin (silk produced by silkworms and spiders)

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Tertiary Structure
Refers to the structure formed by extensive folding of a single polypeptide chain à compact
shape, gives overall 3-D shape of protein
• Bond: 4 types of interaction
o Hydrophobic interaction (weakest)
§ Between non-polar R-grps which interact and cluster at the core of the
protein (mutual exclusion from water)
§ Polypeptide folds to shield hydrophobic R-grps from aqueous
environment (hydrophobic point inwards; hydrophilic face outwards)
o Hydrogen bond
§ Between polar R-groups
§ Electropositive (H of –NH or –OH) and electronegative (O of C=O, N of N-
H) atoms form H bonds
§ Individually H bond is weak; but collectively it is strong
o Ionic bond
§ Between oppositely-charged groups of amino acids
§ COO- (acidic) and NH3+ (basic) found on R groups or at the ends of a
polypeptide chain
§ Change in pH à alter the changes and hence ionic bonds
§ Bonds are weak under aqueous conditions à charged groups are
surrounded by H20 molecules
o Disulphide bond (strongest)
§ Between 2 cysteine amino acid residues brought together by folding of
polypeptide
§ Strong covalent bonds à contribute to toughness (increase in bonds,
increase stability of protein to heat denaturation)
• Steric/spatial relation of residues that are far apart on chain as well as those which are
adjacent

Quarternary structure
• Polypeptide = subunit
• Bonds: ionic boncs, hydrogen bonds, disulfide bonds & hydrophobic interactions
• Constituent chains may be identical or different
• 2 subunits = dimers
• more than 2 subunits = oliomers
• Not all proteins have quarternary structure à E.g. lysozome, myoglobin and albumin

Fibrous protein Globular protein

Long polypeptide chaings forming long Long polypeptide chains folded into
strands spherical shape
Length of polypep/sequence of a.a. may Length of polypep/sequence of a.a. are
very slightly between 2 samples of same always identical between 2 samples of
protein the same protein
Insoluble in H20 à has hydrophobic R Soluble in H20 à have hydrophilic R
groups on the exterior of their molecules groups on exterior of molecules which
interact with H20
Repetitive regular sequence of amino Irregular sequence of amino acids
acids

26
Functions as structural and contractile Functions in a variety of metabolic roles.
proteins (e.g. haemoglobin, enzyme)
COLLAGEN HAEMOGLOBIN
Structural protein; essential component Transport protein; pigment which
of connective tissue in tendons, bone, transports 02 found in rbc
skin and teeth (most abundant fibruous Structure
protein) • Quarternary structure: 4 polypeptide
Structure subunits (2 b-globin + 2 a-globin)
• helical polypeptide chain à • Each subunit = polypeptide
tropocollagen à fibril à fibres component ( globin) +
• 3 helical polypeptide chains prosthetic/non-protein component
wound around each other like a ( haem group)
rope – tropocollagen o Haem group: poryphin ring an
• H bonds further stabilize it and Fe2+ (which binds to O2, so 1 Hb
increase rigidity and tensile can carry up to 4 O2)
strength • Bonds: weak hydrophobic
• each helical polypeptide: about interactions and hydrogen bonds à
1000 a.a. (glycine-X- allows subunits to move, allowing a
proline/hydroprline) change in position that influences its
o glycine (smallest a.a.) à fits affinity for O2
restricted space @ centre of Cooperative binding of O2
triple helix (3 chains come 1 O2 molecule + 1 Hb subunit = structural
together) à tropocollagen change in remaining 3 subunits so that
forms inelastic tight coil their affinity for O2 increases (results in
o proline à bulky/inflexible à rapid uploading of O2 onto other
confers rigidity subunits) and vice versa
• unique secondary structure
• tropocollagen cross-links
(covalent bonds involving lysine
residues) with neighbouring tropo
running parallel à Forms fibrils à
fibrils unite to form fibres
o staggered/overlapping
minimizes points of
weaknesses along length

27
Learning Module No. 6

Topic: Enzymes
Materials: Laptop and Internet Connection
Assessment: MCQ Post-Test via Infinit

Learning Outcomes:
At the end of the session, the students should be able to:
1. Recall the structure and characteristics of enzyme
2. Understand the nomenclature and classification of enzyme
3. Explain the activity of the enzyme
4. List the appropriate methods in quantifying enzymes

Enzymes

Activation energy is the energy required to start a reaction. All chemical reactions need
activation energy. Enzymes reduce the activation energy required to initiate a reaction.

• Enzymes are protein molecules that increase the rate of a chemical reaction (catalyst).
• They reduce the activation energy required to start a reaction
• They participate in reactions but are not used up or permanently altered in the reaction and
are available for reuse
• Without enzymes, metabolism is too slow for life to exist
• Enzymes are specific for one reaction - each enzyme acts on only one substrate. This is
because the active site of an enzyme matches that of only one substrate.

Enzyme: Breaks down:

Amylase Glucose
Maltase Maltose
Protease Protein
Lipase Lipids/fats
Aminopeptidases Peptides (amino acids)

w Many enzymes are intracellular – they are used within cells that produce them
w Some enzymes are extracellular – they are secreted by cells and act outside. Eg: digestive
enzymes.

Exergonic Reactions (Catabolic) Endergonic Reactions (Anabolic)

Ø Break down of substrates Ø Combine substrates
Ø Releases energy Ø Requires energy
Ø Eg: Cellular Respiration Ø Eg: Photosynthesis

Coenzymes and cofactors

Many enzymes require other, non-substrate molecules to be present to increase the ease with
which the enzyme binds to the substrate.

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• Cofactors are non protein parts required by enzymes before they act. These parts include
metallic ions such as zinc and magnesium.
• If a cofactor is an organic molecule it’s called a coenzyme.
• Coenzymes are organic non-substrate molecules required to be present to increase the
ease with which the enzyme binds to the substrate. Many are vitamins, or are derived from
vitamins.

Factors Affecting Enzyme Activity

• The rate of an enzyme reaction is influenced by: pH, temperature, enzyme concentration,
substrate concentration & inhibitors.
Temperature
In any chemical reaction, warming the reactants will increase the rate of reaction due to the
increased rate of interaction between molecules. Similarly, decreasing the temperature will
reduce the movement of molecules, hence the rate of the reaction will slow. Freezing the
reactants will stop the reaction.

§ Enzymes have an optimal temperature, reflecting the optimal temperature of the organism.
§ If too much heat is applied the enzyme will permanently denature - causing it to permanently
lose its 3D shape, resulting in the reaction slowing and then eventually stopping.
§ When the temperature is too low, the enzyme will become inactive (but not denatured). It will
become active again once when it reaches optimal temperature.

pH
§ Enzymes have optimal pH reflecting the pH of its working environment. Mostly pH 7 in
animals.
§ A change in pH from the optimum will change the hydrogen bonding between amino acids.
Hence, it will change the 3D structure of the enzyme and the shape of the active site, affecting
its ability to combine with substrate molecules. When this happens, the rate of metabolic
reaction declines.

Inhibitors
w Competitive inhibition – inhibitors act by binding to the active site of the enzyme.
w Non-competitive inhibition – inhibitors act by binding to a distinct (allosteric) site away
from the active site and changing the shape of the active site

- Competitive inhibitors can be overcome by increasing the concentration of substrate. This

increases the likelihood that a substrate molecule will interact with the active site instead of
an inhibitor molecule.
- Non-competitive inhibitors cannot be overcome this way.

Substrate & product concentration

§ Increasing substrate concentration will increase the rate of the reaction until the enzyme is
saturated. At this point, all enzymes and active sites are constantly occupied.

§ Increasing enzyme concentration will increase the rate of the reaction as there are more
enzymes reacting with substrates.

§ As a reaction progresses, the increasing concentration of product will also begin to exert an
effect, slowing the rate of reaction.

29
Learning Module No. 7

Topic: Nucleic Acid

Materials: Laptop and Internet Connection
Assessment: MCQ Post-Test via Infinit

Learning Outcomes:
At the end of the session, the students should be able to:
1. Recognize the structures of nucleic acids
2. Describe protein synthesis
3. List the appropriate methods in quantifying enzyme

Nucleic acids

Our class on DNA is divided into 3 parts: (I) Genetics (II) DNA structure (III) Concepts and
applications.

I. Genetics: In the primordial period, simple molecules were formed from atoms and from these
molecules, macromolecules were formed. These macromolecules formed life and all living
organisms. The classical genetic and heredity observations in the 19th century started the search
for the origin of life.

The transforming principle of DNA was demonstrated from the experiment in which non-
pathogenic (R-form) and virulent (S-form) but heat treated bacteria, when co-injected, could kill
the mice. After that, the link between genes (DNA) and genotype / phenotype was established.

II. DNA structure: The genomic DNA of a eukaryotic cell is located in a special organelle, the
nucleus, whereas in a prokaryotic cell there is no nucleus. In a virus, including bacteriohage, the
genome is packed efficiently. In a human cell, the complete genetic DNA is organized into 23
pairs of chromosomes.

Chromatid is one of the two identical copies of DNA in a chromosome. The two copies
approach each other at the centromere. The ends of DNA in a chromosome are called telomere.
The location of a gene in a chromosome is marked as, say, 7q31.2 where 7 refers to the
chromosome number, q is the long arm (the short arm of the chromosome is called ‘p’), 3 refers
to the region of a chromosome when colored using a particular process, 1 refers to band 1 in that
region and 2 refers to a sub-band within band 1.

In the chromatin, DNA is wound around the histone core (made by 2 copies each of the
H2A, H2B, H3 and H4 proteins) and clamped by the H1 protein. Anytime this DNA is accessed
for any biochemical reaction, there will be physical rearrangement of DNA and the histone core
and furthermore the histone proteins undergo chemical modifications, like acetylation and
methylation.

Two strands of DNA form duplex DNA through base-pairing. In a basepair, the two bases
are unlikely to be perfectly aligned or coplanar. In the same token, two adjacent basepairs also
need not be perfectly parallel to each other.

30
 There are three forms of DNA: B-DNA, A-DNA and Z-DNA. The B form is the physiological
form. The other two forms are man-made from specific sequences. While the first two forms are
right handed helices, the last one is left-handed. In the B-form, the minor groove is narrow and
the major groove is wide whereas in the A and Z forms, the groove widths are nearly the same.
Also, a basepair in the B-form cuts the helical axis whereas in the A-form, a basepair is very much
away from the helical axis. However, in the Z-form a basepair lies in-between.

Supercoiled DNA: In a chromosome (or even in a circular plasmid), DNA exists in a supercoiled
form. Several studies have established the connection between the number of base-pairs (linking
number, twist) and the level of supercoiling (writhing number). Assume there are 260 B-DNA
base-pairs (10 base-pairs will form one full turn, Fig. 1; start from base-pair 1 on a strand and
come to the same but one earlier position on the same strand after 10 base-pairs; the next 10
base-pairs form the next one round and so on).

Now, convert the linear DNA into circular DNA by

connecting the ends of the same strands. The twist T = total
base-pairs / 10 = 260/10 = 26. The linking number is the
number of times one strand crosses the other, which is also 26.
So the equation becomes,

10 L = T + W; or 26 = 26 + 0
9
Now cut only one strand and unwind that strand two
8
times and reconnect the ends. That means, L becomes 24. In
7 order to balance the above equation, 24 = 26 – 2 or W becomes
6 -2. Or, the new circular adjusts (writhes) with two cross-overs.
5 If you over-wind by two, L = 28 and W = +2. Even now, the
4 circular DNA writhes by 2 but in the opposite direction.
3
2
1
Strand-1 Strand-2
Fig. 1. B-DNA Apart from DNA, RNAs are also very important in
several cellular processes. There are 3 types of RNA, mRNA,
rRNA and tRNA. Of these 3 classes, the tRNA is normally depicted in the ‘clover leaf’ form,
displaying its amino acid acceptor region and the anti-codon region. An amino-acyl tRNA
synthetase enzyme attaches a corresponding amino acid to the tRNA.

III. Applications and concepts: There are several applications and processes that
involve nucleic acids. However, due to limitation of time, we will learn only a few applications.

1. DNA replication: In molecular biology, the important fundamental processes are: the
cell cycle (including DNA replication – the making of DNA using a DNA template), transcription
(the making of mRNA using a DNA template) and translation (the making of a protein using mRNA
as a template). The next level of events includes reverse transcription (the making of DNA using
an RNA template) and the making of RNA using an RNA template. The making of a protein using
a DNA template is not yet known.

31
 In prokaryotic DNA replication, DNA is unwound by enzymes like helicases and long
leading strands ( for the parental 3’ to 5’ strand) and several short lagging strands (for the parental
5’ to 3’ strand) are made by the DNA polymerase. The short fragments are joined by ligases. If
there is any problem during DNA synthesis, like base-pair mismatch, selected enzymes fix those
problems.

In a eukaryotic cell, there are several origins of DNA replication (dedicated sequences in
DNA) in a chromosome. DNA replication must be initiated only once per origin per cell cycle. First,
origin replication protein complex (ORC) binds to the origin of replication. The CDC6 protein
(CDC28 in yeast) binds to ORC. The CDT1 protein binds to CDC6. Next, the mini chromosome
maintenance proteins 2 to 7 (MCM 2-7) bind to the above proteins. The assembly of all these
proteins is called ‘licensing’ and the above complex of all these proteins is called the pre
replication complex (pre-RC).

There are two modes by which DNA re-replication is prevented. The first mode is through
the involvement of cyclin dependent kinases (CDKs). We are not going to review that mode here.
The other mode is through the involvement of geminin, a protein.

Once DNA replication is initiated, Geminin binds to Cdt1 and primes it for degradation.
Once Cdt1 is removed from the pre-RC, there cannot be another DNA replication firing. At the
end of the cell cycle, even geminin is degraded. This way, DNA replication takes place only once
per cell cycle. We have published the structure of geminin. The geminin-Cdt1 complex structure
is also published by another group.

DNA repair: Several types of problems happen during DNA synthesis. One such repair is
DNA mismatch repair.

In a prokaryotic cell, during DNA synthesis, if there is a base-pair mismatch, immediately

DNA synthesis is halted. The MutS protein, as a dimer, first binds at the mismatch site. Then MutL
binds. These proteins pull the sector where the mismatch is present. Next, MutH binds and form
the end clamp. An endonuclease is recruited to cut the mismatch site and DNA is removed from
the region. The DNA polymerase enzyme remakes the stretch again without any mistake. The
process of mistmatch repair is almost the same in a eukaryotic cell.

2. Cloning: In conventional sexual reproduction or in vitro fertilization (IVF), an egg is

impregnated by a sperm cell. But in cloning, the nucleus of an egg is removed and a nucleus from
any suitable cell from an individual is implanted. This cell grows with the same genetic make-up
of the nucleus donor (not the egg donor).

3. DNA microarray: This development is an important tool to study how a normal cell and
an affected cell (say, a cancer cell) behave and what are the genes that are up-regulated and
down-regulated. On a commercial DNA chip, unique and short single stranded DNA fragments of
all known human genes (as of today) are immobilized on glass. Take a normal cell and a cancer
cell. Make complementary DNA for all the RNAs in the cells. Treat the normal cell DNA with a dye
(say green) and that of the cancer cell with a red dye. Now pass the two pools of DNA through
the chip. The genes that are active only in the normal cell (thereby making mRNA and hence
cDNA) will bind to their complementary fragments (immobilized on the chip) and will emit green
signal when detected. Similarly, the genes that are active only in the cancer cell will bind to their
complementary fragments and will emit red signal. The genes that are common to both cells will
give out yellow signal. From this we can learn which genes are upregulated and down regulated
in a particular cell for a particular disease condition.

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 4. CpG methylation and gene silencing: The studies on how a tumor suppressor gene
is silenced in a cancer cell led to the discovery of CG methylation and gene silencing. Statistically,
the occurrence of C, followed by G, in a sequence seems to be very random. However, in most
of the genes, especially in the promoter region, CGs occur more frequently, called the CG islands.
The C of these CG bases get methylated. A set of proteins, called Methylated DNA Binding
Domain (MBD) proteisn (MeCP2, MBD 1-4) bind to these methylated regions of the promoter and
do not allow any access to RNA polymerase, thereby preventing transcription of the gene. The
pattern of CG methylation in a normal cell and cancer cell can identify the level of gene silencing
(and hence the indirect clues on the level of disease progress and treatment). CG methylation
can be studied using a technique called pyrosequencing.

5. Transgenic / reporter genes: Selected color displaying proteins, like green fluorescent
protein (GFP), can be used as reporters to identify the location of protein expression for a protein
of interest. The GFP gene is attached to the gene of our interest and injected in an embryo and
the location of protein expression is visually observed. Such techniques can be used to generate
multicolored ornamental fish for the same species.

Gene therapy: Certain diseases, like insulin dependent diabetes and hemophilia, occur
because of the lack of related proteins. The normal mode of treatment is administering these
proteins by injection. However, in gene therapy, in the case of diabetes, the gene of good insulin
is first inserted into a suitable vector with a promoter. This construct is delivered to the organ of
expression, say pancreas, using a modified virus, which is used as a delivering agent. Similarly,
the gene of Factor VIII is delivered to the liver. These genes express the corresponding proteins
at the corresponding organ.

6. DNA protein interaction: Several proteins interact with DNA. For example,
transcription factors bind to the promoter / enhancer regions of a gene. Restriction enzymes bind
to and cut DNA. DNA polymerase is involved in DNA replication and RNA polymerase is important
for transcription. Furthermore, amino-acyl tRNA synthetases bind to tRNAs and attach
corresponding amino acids to them.

7. RNA interference: Most of the free forms of RNA, messenger RNA molecules in
particular, are single strands. tRNAs and selected RNA regions are double-stranded. Many
viruses, however, form long stretches of double-stranded RNA when they replicate.

When our cells find double-stranded RNA, it is often a sign of an infection. However, plant
and animal cells have a more targeted defense that attacks the viral double stranded RNA directly,
termed RNA interference.

Viral double-stranded RNA are cut into pieces (about 21 base-pairs), called small
interfering RNA (SiRNA) by the protein Dicer. The argonaute protein strips away one strand from
the siRNA, and then looks for any messenger RNA that matches it. If it finds some, it cleaves the
RNA, destroying it. In this way, the cell removes any messenger RNA that is the same as the
original double-stranded piece found and processed by dicer.

Based on this principle, we can synthesize a non-natural interfering RNA, then insert it
into a cell to destroy any messenger RNA that we desire. Researchers use these small RNA
molecules to fight disease, for instance, using them to knock out cancer genes.

33
 8. RNA modifying enzymes: RNA has to be modified in selected cellular processes. For
example, uridine is modified to pseudo-uridine by pseudo-uridine synthase enzymes. Another
example is PARN, which truncates the poly-A tail of mRNAS.

34