PROTEINS AND AMINO

ACIDS

1
 FUNCTIONS OF PROTEINS
Proteins perform several functions in living system
1. They are responsible for strength and structure of body such proteins are known
as structural proteins. (example elastin and collagen)

2. The biochemical reactions taking place in the body are catalyzed by proteins
called enzymes (which participate in chemical reaction and makes the process
fast).

3. Proteins are responsible for transportation of metabolites known as transport

proteins. (also responsible for respiration)

4. Proteins which regulate metabolic pathway are known as regulatory proteins.

(example: insulin)

5. Proteins protect the body from infection and other toxins such proteins are
known as defense proteins (Example: antibodies, immunoglobulins)

6. Some proteins are required for mechanical work and are known as muscle
proteins. 2
 AMINO ACIDS
• Amino acids are group of organic compounds containing two functional groups
a) Amino (-NH2)
b) Carboxyl (-COOH)
• These functional groups are attached to Alpha- carbon and therefore are called ɑ-
amino acids.
• The general structure of amino acid is given below.
• There are more than 300 amino acids in nature but out of these only 20 are
known as standard amino acids.
• The genetic code incorporates only 20 amino acids to synthesis proteins.
• In the structure given below R represents side chain attached to alfa carbon and
each of 20 amino acids have different R group or side chain i.e all the amino
acids differ in structure of side chain.

3
 AMINO ACIDS

Based on Essential Non essential

structure Based on amino acids amino acids
-Aliphatic amino polarity
acids -Non polar amino
-Aromatic amino acid acids
-Amino acid -Polar amino acids
containing hydroxyl
group
-Sulfur containing
amino acid
-Acidic amino acids
and their amides
-Basic amino acids
-Imino acids

4
 CLASSIFICATION OF AMINO ACIDS BASED ON STRUCTURE

A. Amino acids with aliphatic side chain

5
B. Amino acids containing hydroxyl group (-OH)

6
C. Sulphur containing amino acids

7
D. Amino acids and their amides

8
E. Basic amino acids

9
F. Aromatic amino acids

10
G. Imino acid

11
 CLASSIFICATION OF AMINO ACIDS BASED ON POLARITY

1. Non polar amino acids

• These amino acids are also known as hydrophobic or water hating.
• They have no charge on the R group.
• Example: alanine, valine, leucine.

2. Polar amino acids

• These amino acids are also known as hydrophilic or water loving.
• They have no charge on R group.
• They possess hydroxyl, amide or sulfhydryl group which participate in
hydrogen bonding.
• Example: glycine, serine, cysteine.

12
 CLASSIFICATION OF AMINO ACID – ESSENTIAL AND NON ESSENTIAL

1. Essential or indispensible amino acids

• These are amino acids which cannot be synthesized in the body and therefore be
supplied through the diet.
• They are known as essential amino acids.
• Required for proper growth and maintenance.
• Example:This group contains 10 amino acids
Arginine, valine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine,
threonine, tryptophan

2. Non essential amino acids

• These are amino acids which are synthesized in body.
• They need not be consumed in diet.
• The body can synthesize 10 amino acids.
• Example: glycine, alanine, serine, cysteine, aspartate, asparagine, glutamate,
glutamine, tyrosine, proline

13
 ESSENTIAL AMINO ACIDS NONESSENTIAL AMINO ACIDS

Valine Glycine

leucine Alanine

Isoleucine Tyrosine

Phenylalanine Aspartic acid

Tryptophan Glutamic acid

Lysine Aspargine

Arginine Glutamine

Histidine Cysteine

Methionine Serine

Threonine Proline

14
 PROPERTIES OF AMINO ACIDS
Physical properties of amino acids
 Solubility
• All amino acids are soluble in water but the extent to which they are soluble
varies.
• R group determines the solubility.
• Polar amino acids are highly soluble in water.
• Non polar amino acids are soluble in organic solvents (chloroform).

 Melting point
• As they are high molecular weight compounds they have higher melting point.
Mostly above 200.

 Taste
• Amino acids can be sweet(glycine), bitter(arginine) or tasteless(leucine)

15
 Optical activity
• For the compound to be optically active it should have an asymmetric carbon,
which means carbon attached to four different groups.
• All amino acids except glycine are optically active due to asymmetric carbon.
• Therefore amino acids exists as stereo isomers (non superimposable mirror
images of each other).

Alanine- carbon
atom is attached to
four different groups

 Amino acids as amphoteric molecules or ampholytes

• Amino acids contain acidic (-COOH) group and basic (-NH2) group therefore it
can act as acid by donating proton and as a base by accepting proton.
• Hence amino acids are known as amphoteric molecules or ampholytes.
16
 Zwitterion or dipole ion
• The term zwitter means hybrid.
• Amino acid can be defined as hybrid molecule consisting of positive as well as
negative charge ionic group or charge.
• When present in solution it exits as zwitterion form i.e. having both positive and
negative charge.
• In acidic solution it has positive charge and in alkaline solution it has negative
charge.
• At one point (pH) in the solution it has both positive and negative charge and
exists as zwitterion.

17
In acidic solution or at extreme acidic pH it has positive charge and in alkaline
solution or extreme basic pH it has negative charge. At one isoelectric pH it acts
as zwitterion having both positive ang negative charge.
18
 Isoelectric pH
• For any amino acid there exist an intermediate pH at which the net charge
present is zero, this pH is known as isoelectric pH of amino acids.
• At this pH amino exists at zwitterion form.
• At isoelectric pH they are very insoluble in solution and therefore precipitate
out.
• This property is used to isolate amino acid from mixture of amino acids.
• Each amino acid has different isoelectric pH
Example:
Aspartic acid-2.87
Glutamic acid-3.22
Glycine-5.97

19
 COVALENT INTERACTION

Peptide bonds
• Amino acids in proteins are joined together by peptide bonds.
• The bond is formed between alfa-carboxyl and alfa-amino groups of different
amino acids.

20
 NON-COVALENT INTERACTION

In addition to covalent bonds there are also non covalent bonds in amino acids.
Like hydrogen bonds, hydrophobic bonds and ionic bonds.

Hydrogen bonds
The oxygen atom of carbonyl, hydroxyl and carboxyl forms a weak bond with
hydrogen atom

hydrogen bond.

Ionic bonds
Amino acids which have ionic side chains form ionic bonds.
That is bond between +amino group and –ve (-COOH) group.

21
Hydrophobic bond
Amino acids having non polar side chain are responsible for such interaction.
The non polar group come close to each other and form a bond.

22
 INTRODUCTION

• Proteins are most abundant organic molecules of living system.

• Proteins are naturally occurring polymers made up of amino acids and linked
together by peptide bonds.

• Single unit of amino acid is known as monomer, when many monomers combine
together they form polymers.

• Proteins are made from 20 ɑ-amino acids.

• On hydrolysis proteins give amino acids.

• Therefore to understand proteins we first study the structure of amino acids.

• They have a high molecular weight and are found in living systems.
23
 CLASSIFICATION OF PROTEINS

I. Functional classification of proteins (based on function)

This classification is based on functions they perform.
1. Structural proteins-Keratin of hair and nails, collagen of bones.
2. Enzymes or catalytic proteins-Pepsin
3. Transport proteins -Hemoglobin, serum, albumin.
4. Hormonal proteins -Insulin, growth hormone.
5. Contractile protein-Actin, myosin
6. Storage protein- glutein,ovalbumin.
7. Genetic protein- nucleoprotein
8. Defense proteins- snake venom, immunoglobulins.
9. Receptor proteins- hormones and viruses.

24
II. Protein classification based on chemical nature and solubility
A. Simple proteins
These contain only amino acid residues.
They are further classified into.
 Globular proteins
Water soluble, oval in shape and are digestible.
• Protamines
These are basic proteins of low molecular weight (3000-10000). They are not heat
coagulable. (do not become solid on heating)
• Histones
These are basic proteins with higher molecular weight. They are heat coagulable.
Widely distributed in body.
• Albumins and globulins
These are globular proteins found in cells.
• Prolamines
Found in wheat (gliadin)
• Globins
25
These are found along with histones.
 Scleroproteins
Has fiber like shape, insoluble in water, and non digestible.
• Collagens
They are connective tissue proteins and on boiling with water become soluble and
digestible.
• Elastins
These are found in arteris.
• keratins
These are proteins of skin, hair and nails.

26
B. Conjugated protein
Along with amino acid these protein contain non-amino acid moiety.
The non-amino acid part is known as prosthetic group or conjugating group.
Other substance apart from amino acid impart characteristic properties.
They are further classified into
• Nucleoproteins
These are formed by combination of histone with RNA or DNA.
• Chromo-proteins
These are soluble proteins combined with chromophoric (colored) group.
The colored group is haem and riboflavin.
• Phosphoproteins
Along with amino acid it contains phosphate group. Milk casein is an example.
• Glycoproteins
Consists of amino sugars (carbohydrate and amino acids), sulphate and sugar acids.
• Lipoproteins
Combination of proteins with lipids. Found in brain and membrane.
• Metalloprotein
Various metals (Fe, Co, Zn) are attached to simple proteins. 27
C. Derived proteins
These are degraded or denatured products of simple or conjugated proteins.
They are classified as follows
 Primary derived protein
• Coagulated proteins
These are denatured proteins produced by heat, acids or alkalies.
Example: albumin (egg white)
• Proteans
These are the 1st or the earliest products of protein hydrolysis by enzymes, dil acids,
alkalies.
Example: fibrin from fibrinogen
• Metaproteins
These are 2nd products of protein hydrolysis.
In this stronger acids are used.
Example: acid and alkali metaproteins.
 Secondary derived proteins
These are progressive hydrolysis products of proteins.
Example: polypeptides, peptides etc. 28
29
III. Nutritional classification of proteins (based on nutritional value)
• Complete proteins
These proteins have all 10 essential amino acids required for human body for
growth.
Example: egg albumin, milk casein.
• Partially complete proteins
They partially lack one or more essential amino acids, hence promote moderate
growth.
Example: wheat and rice proteins
• Incomplete protein
These proteins completely lacks one or more amino acids.
They do not promote growth at all.
Example: gelatin

30
 PROTEINS AND ITS STRUCTURE

• Proteins are polymers of L-alpha-amino acids.

• Proteins have a complex structure and can be divided into four.
• The term protein is used for polypeptides containing more than 50 amino acids.

a) Primary structure
• It is a linear structure of protein and forms the backbone of proteins.
b) Secondary structure
• It is the arrangement of protein structure in space(spatial arrangement) by
twisting the polypeptide chain.
c) Tertiary structure
• It is the three dimensional structure of proteins.
d) Quaternary structure
• Composed of two or more polypeptide chains known as subunits the spatial
arrangements of these subunits is known as quaternary structure.

31
32
 PRIMARY STRUCTURE OF PROTEIN
• Each protein has a unique sequence of amino acids.
• The primary structure of proteins is responsible for functions.
• Mostly many diseases are caused due to abnormal sequencing of amino acids.
• Amino acids are held together by peptide bonds to form polypeptides or
proteins.
• When the amino group of amino acid combines with carboxyl group of another
amino acid the bond is known as peptide bond.

33
34
 SECONDARY STRUCTURE OF PROTEINS
The conformation of polypeptide chain by twisting and folding is referred to as
secondary structure.
There are two types of secondary structure
1. ɑ-helix (alpha-helix)
2. ß-sheet (beta-sheet)

 Alpha-helix
• It is the most common spiral structure of protein.
• It has rigid arrangement of polypeptide chain.
• It is tightly packed coiled structure with amino acid side chains.
• The structure of alpha-helix is stabilized by hydrogen bonding (i.e hydrogen bond
is formed)
• The hydrogen bond is formed between H atom attached to peptide peptide N and
O atom attached to peptide C
• Each hydrogen bond is weak but combination of all hydrogen bond is strong.
• All the peptide bonds except the first and last participate in hydrogen bonding.
35
• Each turn of the coil contains 3.6 amino acids and the distance travelled is 0.54
nm. The distance between two amino acids is 0.15.
• The formation of alfa helix requirs lowest energy.
• The right handed alfa-helix is more stable than left handed.
• Acids, base etc interfere with structure of alfa-helix.

36
Alfa-helix

37
 Beta-sheet or beta-pleated sheet
• It contains extended polypeptides.
• In this the hydrogen bonds are formed between the neighboring segments of
polypeptides.
• The peptide is held together giving a sheet like structure.
• Can be parallel (same direction) or antiparallel(opposite direction).

38
 TERTIARY STRUCTURE OF PROTEINS
• It is the three dimensional arrangement of protein structure.
• In this the hydrophobic side chain is held inside and hydrophilic groups are held
outside (surface).
• The above arrangement gives stability to the molecule.
• The interactions include
Hydrogen bonds
Hydrophobic bonds
Ionic bonds
Disulphide bonds

39
 QUATERNARY STRUCTURE OF PROTEINS
• Some proteins contain 2 or more polypeptides held together by non-
covalent bonds.
• The two polypeptides can be identical or unrelated.
• These 2 polypeptide chains are called oligomers and single polypeptide is
called monomer.

Protein with 4 polypeptides

40
 POLYPEPTIDES
• The condensation product of amino acid is peptide.
• Peptide refers to condensation of 2-20 amino acids.
• Polypeptides contain 20-50 amino acids.
• Proteins contain more than 50 amino acids.
Example: oxytocin and vasopressin are peptides.

 Oxytocin
It is hormone secreted by pituitary gland.
Contains 9 amino acids.
 Vassopressin
It is present in pituitary gland.
It stimulates kidney to retain water.
 Aspartame
It is a dipeptide containing aspartic acid and phenylalanine.
It has low calorie and is 200 times sweeter than sucrose.
Used as artificial sweetner. 41
 PROPERTIES OF PROTEINS
 Solubility
Proteins have huge size therefore form a colloidal solution in water.

 Molecular weight
The molecular weight of proteins varies depending upon number on amino acids.
They have high molecular weight. Mol weight ranges from 40-40,000.

 Shape
Their shape varies.
Insulin has globular shape, albumin has oval shape, fibrinogen has elongated shape.

 Isoelectric pH and zwitterion.

Same as amino acids.

42
 Precipitation of proteins
Proteins get dehydrated when salts such as ammonium sulfate are added.
Dehydration also results by addition of salts of heavy metals like Pb2+, Zn2+ etc.
Alcohol also dehydrates protein.
Tannic acid and picric acid also dehydrates protein.
Dehydration of protein results in precipitation of proteins.

43
 Denaturation of proteins
• The change in structure of proteins is called denaturation of proteins.
• This results in loss of secondary, tertiary, quaternary structure of proteins.
• Due to this there is change in physical, chemical and biological properties of
protein.
• Physical agents which cause denaturation of proteins are- heat, UV rays, Violent
shaking etc.
• Chemical agents which cause denaturation of proteins are- acids, alkalies,
organic solvents.
Characteristic of denaturation
• The helical structure of protein is lost.
• Primary structure of protein remain intact.
• It looses it biological activity.
• The solubility pattern changes.
• Viscosity increases.
• More easily digested.
• Denaturation is mostly irreversible.
• Results in coagulation or flocculation. 44
 REACTIONS OF PROTEINS

• Heat test
When protein solution is heated in boiling water bath, it gets coagulated and loose
their biological activity.
This is called denaturation of protein.
As heat causes the denaturation, it is called thermal denaturation.

• Test with trichloroacetic acid (TCA)

TCA is used to precipitate proteins from their solution.
Due to addition of acid proteins get denatured.

• Biuret test
Biuret reagent contains copper sulphate in alkaline solution.
Also contains sodium potassium tartarate.
Proteins reduce the cu2+ ions to cu+ ions.
When proteins are treated with biuret regent it gives violet color.
This test is used for identification of amino acids. 45
• Hydrolysis test
Proteins on hydrolysis gives amino acids.
Hydrolysis can be carried out by acids like hydrochloric acid , conc. Sulphuric acid
etc.
• Xanthoproteic reaction
Same as amino acids.
Nitration of aromatic amino acids will give yellow color which on addition of
alkali turns orange.
• Millons test
Same as amino acids
Proteins react with mercuric sulphate in presence of sodium nitrite and sulphuric
acid to give red color.
• Precipitation test
Proteins are precipitated by using different reagents like
1. Salts- Example: ammonium sulphate
2. Organic solvent- Example: alcohol
3. Heavy metals-Example: copper or mercury.
4. Acids- Example: TCA, acetic acid or hydrochloric acid. 46
 DISORDERS OF PROTEIN DEFICIENCY

• Deficiency of protein from early childhood is regarded as disease.

1. Kwashiorkor
It is a severe protein malnutrition disease.
Appears in age of 1-4 years.
Symptoms
Retarted growth
Edema (excess water in the body tissues which results in swelling of body)
Alterations in skin. (patchs or redness on skin)
Hair pigmentation and texture changes. (which means color of the skin becomes
dark or changes)
Liver enlargement
Vomiting and diarrhoea and stools contain undigested food.

47
Causes
Poor maternal (mothers) health.
Large family size
Termination or delayed of breast feeding.
Environmental conditions.
Over diluted cows milk

Cure
Diet rich in protein such as milk and eggs.
Soya beans are best vegetable source of protein.

48
2. Marasmus
This disease is same as kwashiorkor but occurs in infants i.e below 1 year of age.
Causes
Due to nutritional deficiency in carbohydrates, proteins etc.
Cure
Providing diet rich in proteins.

3. Nutritional oedema
It is a swelling caused due to insufficient protein intake.
Causes
Long continuous deprivation of proteins in diet.
This defeciency in adults is rare.

49
Symptoms in adults
Loss of weight
Anemia
Constant infection
Frequent loose stools
Lethargy
Delay in healing wounds
Cure
Diet rich in proteins
Soyabean, milk and eggs in diet.

50