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Chemistry of Proteins
Definition:
Proteins are organic compounds with a
high molecular weight formed of
carbon, oxygen, hydrogen and nitrogen
and may also contain sulfur,
phosphorus coloring non-protein
organic groups and metal ions.
They are polymers formed of subunits
called amino acids linked together by
peptide linkage.
Amino Acids
Amino acids are organic acids that contain NH2
group.
They are the structural units of proteins and are
obtained from them by hydrolysis.
The general formula of any amino acid is as
follows:
O
H2N
CH
OH
I-Chemical classification
(According to number of carboxyl and
amino groups)
Amino acids can be classified into:
a) Neutral amino acids.
b) Acidic amino acids.
c) Basic amino acids.
OH
H2N CH
OH
CH3
Glycine
H2N CH
CH CH3
Alanine
CH3
O
O
H2N
CH
OH
H2N
CH
CH2
CH CH3
CH CH3
CH2
CH3
CH3
Leucine
OH
Isoleucine
OH
Valine
O
H2N CH
C OH
H2N CH
C OH
CH2
CH OH
OH
CH3
Serine
Threonine
O
H2N
CH
OH
CH2
Phenyl alanine
H2N
CH
CH2
OH
Tyrosine
OH
Cysteine
S
CH2
H2N CH C OH
CystineO
O
H2N CH C OH
CH2
CH2
S
CH3
Methionine
O
C OH
CH2
N
NH
imidazole group
Histidine
H2N CH
O
C OH
CH2
HN
indole group
Tryptophan
O
C OH
CH
HN
CH2
CH2
CH2
Proline
O
C OH
CH
HN
CH2
CH2
CHOH
Hydroxyproline
C O
OH
Glutamic acid
O
H2N CH C OH
CH2
CH2
C O
NH2
O
H2N CH C OH
CH2
C O
amide
OH
group Asparatic acid
Glutamine
O
H2N CH C OH
CH2
amide
NH2 group
Asparagine
C O
CH
CH2
OH
H2N
CH
O
OH
CH2
H2N
CH
CH2
CH2
CH2
CH2
NH
NH
NH2
CH2
Ornithine
CH2
NH2
urido group
Citrulline
OH
CH2
NH2
NH
guanido group
Arginine
CH
CH2
CH2
CH2
CH2
NH2
Lysine
OH
H2N
CH
O
C
CH2
CH2
CHOH
CH2
OH
NH2
Hydroxy lysine
II-Metabolic classification:
classification
Amino acids may be classified into
A-glucogenic amino acids, i.e., those
which can be converted into glucose,
B-ketogenic amino acids, i.e., those
which can be converted into ketone
bodies
C-mixed amino acids, i.e., those which
can be converted into both glucose and
ketone bodies.
Ketogenic
Leucine
Glucogenic
Lysine
Isoleucine
Tyrosine
Tryptophan
Phenyl alanine
Valine
Leucine
Isoleucine Threonine
Lysine
Tryptophan
Methionine Phenylalanine
Arginine
Histidine
Structure of Proteins
There are 4 levels or orders of
organization of the structure protein
molecule: primary, secondary, tertiary
and quaternary structures.
This complication gives the molecule
its functional domain to explain its
structure-function requirements that if
changes due to mutation will give nonfunctional protein and, therefore, a
disease.
1. Primary structure:
Primary structure is the linear form of
the polypeptide illustrating the total
number, chemical nature, and linear
order of all of the amino acid residues
in the polypeptide chain or chains of a
protein and position of disulfide bonds
if present.
The peptide bonds (primary bond) are
responsible for the primary structure.
2. Secondary structure:
It is the fine folding of polypeptide chain into
specific regular coiled structure or irregular
random coiling held together by hydrogen,
ionic and disulfide bonds.
It is due to the interaction of amino acids
located very close to each other.
3. Tertiary structure:
structure
It is the final three-dimensional form due to
the more complicated course folding and
super-folding of the polypeptide chain in its
secondary level into globular or fibrous form
of different size.
It is due to interaction of amino acids located
far apart (away from each other).
It is the biologically active conformation of
the polypeptide and therefore, is the most
liable to denaturation.
The bond stabilizing the tertiary structure are
disulfide bonds, hydrogen bonds and ionic
bonds
4. Quaternary structures:
structures
Proteins consist of two or more polypeptide
chains in their tertiary structure united by
forces other than peptide bonds are said to
possess a quaternary structure.
Quaternary structure therefore, is the
positional relationship between individual
polypeptides associating to form one protein
molecule.
The bonds responsible for the quaternary
structure are not disulphid or peptide bonds
Subunit
-Sheet
-helix
Random coils
-helix
-Sheet
Random coils
Subunit
Quaternary protein structure
Classification of Proteins
I. According to the biological importance of the
protein:
1) Proteins of high biological value:
These are all proteins of animal origin (with a
few exceptions) and some proteins of plant
origin that contain all the 10 essential amino
acids in well balanced amounts and are
easily digestible.
Examples of animal proteins include; milks
and its products, egg, liver, fishes, red and
while meats.
Examples of the few plant proteins of high
biological value are lentils and broad beans.
1. Fibrous proteins:
They have an axial ratio of more than 10.
Axial ratio = Length/Width of the protein
molecule. They are fairly stable proteins.
Examples,
a. Keratin proteins in hairs, wool, skin, and
most cells. In its native state, it is present in
the form of coiled polypeptide chains called
-keratin. It can be stretched by
denaturation forming -keratin.
b. Myosin is the major protein of muscles.
During muscle relaxation it is called myosin but during muscle contraction, it
undergoes a change in its structure and it
becomes -myosin
2. Globular proteins:
proteins
Their axial ratio is less than 10.
Their peptide chains are folded or
coiled on themselves in a very compact
manner.
They are less stable than fibrous
proteins. Examples are albumin,
globulins, and insulin
A) Simple Proteins
These are proteins which on hydrolysis
produce amino acids only.
Simple proteins are subdivided according to
their physical properties, solubility,
molecular weight and amino acid
composition into:
1-Albumin and globulins .
2-Basic proteins (Histones & Protamines).
3-Acidic proteins (Glutelins & Gliadins ).
4-Scleroproteins(Keratin&Collagen&Elastin)
Albumin
Globulin
- Coagulated by heat.
- Easily digested.
- M.W.: 68 KDa.
- It functions as transporting
protein for elements, vitamins, and
hormones other than keeping blood
osmosis.
3-Acidic proteins
A-Gliadins or Prolamines:
B-Glutelins:
They are plant proteins.
They are soluble in diluted acids but
not in water or diluted salt solutions.
They are very rich in glutamic acid.
They have very large molecular weight
and are heat coagulable.
Examples are oryzenin of rice and
glutelin of wheat.
4-Scleroproteins (Albuminoids)
Scleroproteins are characterized by their
extreme insolubility in water, dilute acids and
the most common reagents.
They are strong fibrous structural proteins
that are rich in sulfur containing amino acids
and hence disulfide bonds.
Their main function is the protection of the
body.
Hairs, nail and connective tissues, contain
scleroproteins and are never present in
plants. The main important groups of
scleroproteins are:
1. Elastins.
2. Collagens.
3. Keratin
A-Elastins:
They are present in the yellow fibers of the
connective tissues in lungs, uterine wall
during pregnancy, tendons and ligaments.
Elastins are also present in the elastic
tissues of tendons and big arteries.
It is rich in alanine, leucine, valine and
proline but deficient in cysteine, methionine,
lysine and histidine.
Boiling scleroproteins with strong acids or
strong alkalis or their digestion by elastase
leads to their hydrolysis to free amino acids
B-Collagens:
Gelatins:
It is the product of prolonged boiling of collagen in
water.
It is easily digested and has the property of
forming a gel on cooling (gel formation).
Gelatin is a very good diet for patients because it
is an appetizer and easily digested.
Gelatin is deficient in certain essential amino
acids. So it is not an adequate protein diet, as it is
deficient in tryptophan and cysteine and contains
very small amounts of methionine (protein of low
biological value).
C-Keratins
Keratins are highly insoluble compounds. They
are insoluble in all protein solvents, and are not
digestible by proteolytic enzymes (e.g pepsin and
trypsin).
Keratins are hydrolyzed by prolonged boiling
with alkalis.
The sulfur content of keratin is high. It is present
in the form of cystine, which is responsible for the
stability and insolubility of keratins.
Most keratins yield histidine, lysine and arginine
amino acids on hydrolysis
B) Conjugated proteins
On hydrolysis, they give amino acids and
prosthetic group (i.e., a non-protein group).
They include:
1. Phosphoproteins:
These are proteins conjugated with
phosphate. Phosphate is attached to
OH group of serine, tyrosine or
threonine present in protein.
They are found in:
a. Casein: milk protein.
b. Vitellin: Egg yolk protein.
2. Lipoproteins:
These are proteins conjugated with lipids
converting them into water soluble
substances. Present in blood, brain and egg.
Cell membrane is of lipoprotein.
Examples: Plasma lipoproteins: see lipid
chemistry.
3. Glycoproteins:
These are proteins conjugated with
carbohydrates in varying amounts attached
as short or long chains.
Examples: Mucous secretion of
gastrointestinal tract and Glycoproteins of
cell wall.
4. Metalloproteins:
These are proteins conjugated with
metals such as;
Iron: e.g., ferritin is intracellular ironbinding protein and Transferrin is an
iron-binding transport protein in the
blood.
Zinc: e.g., Insulin hormone present in
crystals containing zinc.
Copper: e.g., Ceruloplasmin: is a
protein present in blood. It is
responsible for the oxidation of Fe2+
ions to Fe3+ ions.
5. Chromoproteins:
These are proteins conjugated with colored
pigment.
Example; Hemoglobin and cytochrome enzyme
present in mitochondria contains haem pigment,
which is red in color.
6. Nucleoproteins:
These are proteins (protamines or histones)
conjugated with nucleic acids (DNA or RNA).
Examples; Chromosomes: These are proteins
conjugated with DNA.
Ribosomes: They are proteins conjugated with
RNA.
RNA
C) Derived Proteins
They include:
A- Denatured protein: e.g., coagulated albumin
or globulin.
B. Hydrolytic product of protein: e.g.,
H2N CH C HN CH C OH
+ H2N CH C OH
an amino acid
an amino acid
H2O
Dipeptide
HN
CH
O
R
CH2
HN
CH
peptide chain
disulfide bond
S
CH2
CH2
HN
two cysteine
residues
SH
R
CH2
2H
SH
CH
C
O
HN
CH
C
O
peptide chain
CH
peptide chain
hydrogen bond
H
CH
R
peptide chain
2. Hydrophobic bonds:
The non polar side chains of neutral amino
acids tend to associate in hidden core of
protein molecule away from solvent.
O
HN CH C HN CH C
R
HN CH
R
HN CH C
O
3. Electrostatic bonds:
These are salt bonds formed between
oppositely charged groups in the side chains
of amino acids e.g. -amino group of lysine
and the carboxyl group of asparatic acid.
O
H2N
Lysine
CH
O
OH
H2N
CH
CH2
CH2
CH2
CH2
CH2
NH3+
O-
Electrostatic
attraction
OH
Asparatic acid
Denaturation of Proteins
Definition:
It is the loss of the native form of the protein
leading to disruption of its secondary,
tertiary and quaternary structure with the
changes in their physical and chemical
characteristics and loss of their biological
activity.
Causes of Denaturation
Physical causes:
As shaking (mechanical effect), high
temperature, X-rays and atomic
radiations.
Chemical causes:
As organic solvents (e.g. acetone), strong
alkalis and acids and agents that irreversibly
precipitate proteins.
Effects of Denaturation
Physical changes:
changes Increase in viscosity,
decreased solubility and decreased
diffusiblility.
Chemical changes: leads to loss of hydrogen,
hydrophobic and electrostatic bonds but
not peptide and disulfide bonds.The result
are loss of secondary, tertiary and quaternary
structures but not of the primary structure.
Biological changes: which include loss of
enzymatic, hormonal and other biological
properties of proteins. Denatured proteins
(e.g cooking), are easily digested than native
proteins.
Protein Separation
Protein separation is based on
1) Protein solubility
2) Size of protein molecule
3) Charge of the molecule
Methods of protein separation
Chromatography.
Salting out
Electrophoresis.
Dialysis.
Ultracentrifugation.
1- Salting out
It depends on elemination of water from the solution. e.g.
albumin by full saturation with Amm. sulphate
globulin by half saturation with Amm. Sulphate
2-Chromatography
Chromatography is a group of separation techniques, where a
mixture of molecules is separated.
The separated molecules are divided between a stationary
sold phase and liquid mobile phase.
The separation process depends on the tendency of one
type of molecules in the mixture to associate more strongly
with one phase than the other.
3- Electrophoresis
It is movement of charged particles in an
electric field towards the oppositely charged
electrode.
By electrophoresis a mixture of amino acids,
polypeptides or proteins can be separated
into distinct bands by using electric current.
1-Globulin
Albumin
-Globulin
2-Globulin
-Globulin
Fibrinogen
Origin
4-Dialysis
Dialysis means separation of colloids from
crystalloids. Proteins have a high molecular weight
that forms a colloidal solution.
If there is a mixture of proteins (colloids) and salts
(crystalloids) they can be separated by dialysis, i.e.,
by using a semi-permeable membrane. Crystalloids
with very small molecular weight can pass through
this membrane, while colloids can not due to the
large size of their partic
Dialysis membrane
Crystalloids
Colloids
5-Ultracentrifugation:
Using high speed centrifuge, a mixture
of proteins is separated into different
fractions according to their densities.
Lipoproteins can be separated by
ultracenrifugation to chylomicrons,
VLDL, LDL, and HDL.