PROTEINS

Catalysis of Biochemical Reactions

Composition and structure

-Proteins are extremely complex
organic compounds containing carbon,
hydrogen, oxygen,nitrogen and with
few exceptions, sulfur. Most proteins
also contain phosphorus and
specialized protein contains very small
amounts of iron, copper, and other
inorganic elements.
-The presence of nitrogen
distinguishes protein from
carbohydrates and fats. Proteins
contain an average of 16% nitrogen.

ENZYMES
-Enzymes are proteins that catalyze
chemical reactions without being used
up or destroyed in the process
-Used in digestion, releasing of
energy from nutrients for fuel,
triggering reactions that build muscle
and tissue

Functions
They are also involved in the following:
-regulation of metabolic processes

Defense Against Infection

-catalysis of biochemical reactions

( enzymes)

-The Immune Response is a series of

steps your body takes to mount an
attack against invaders

-transportation of
oxygen( hemoglobin)
-defense against infection( antibodies)
-transmission of heredity
characteristics (nucleoprotein)
-muscular activity (contraction)

-Antibody (immunoglobin)-large Yshaped protein produced by Bcells that is used by the immune
system to identify and neutralize
foreign objects such
as bacteria and viruses

-are components of skin , nails as well

as connecting and supporting tissue.
According to the biological role,
we can classify proteins as:
A.Enzymes: catalysts that
specifically interact with the
reactants in a particular reaction
and increase the reaction rate as
much as 1016.

ACID-BASE BALANCE

-Proteins help to maintain a stable pH

level in our body fluid by picking up
extra hydrogen ions when conditions
are acidic, and donating hydrogen ions
when conditions are alkaline

D. Storage proteins: Storage

proteins provide reservoirs of essential
nutrients:

-Otherwise, the resulting conditions of

acidosis or alkalosis could lead to
coma or death

-Ferritin is a iron-binding protein; it

can carry as many as 4,500 atoms of
iron per molecule. 8

-Ovalbumin (egg white) and casein

(milk) are proteins that serve as
reservoirs of amino acids.

E. Contractile and motile proteins:

The proteins that allow cell and tissue
movement, such as
-Actin and myosin in muscle.
B. Regulatory proteins:
Hormones: proteins, such as insulin,
glucagons and etc., that regulate the
metabolism of cells.

-Tubulin in the microtubules of the

mitotic spindle or the eukaryotic
cilium.
F. Structural proteins:

Transcription factors: proteins

that regulate gene expression. Some
repress gene expression (such as lac
repressor, which binds to its target
gene and prevents its transcription);
others activate gene expression (such
as CAP catabolite activator protein)
by binding to target genes and
activating their transcription.

--keratin is the structural protein of

hair, horns, and fingernails.

C. Transport proteins:

-Actin and myosin in muscle.

-Hemoglobin transports O2 in the

blood.

-Tubulin in the microtubules of the

mitotic spindle or the eukaryotic
cilium.

-Serum albumin transports fatty acids

in the blood.
-Membrane transport proteins
transport substances across cell
membranes. Example: the glucose
transporter

Collagen is the structural protein of

bone & connective tissue
Muscular Activity
-proteins that allow cell and tissue
movement

-proteins have very high molecular

masses. A proteins is made up of aamino acids joined by means of
peptide linkage
--5,000 to 8,000,000 molecular
weight
G. Protective proteins:
-Immunoglobulins or antibodies are
the proteins of the immune response.
-Blood-clotting proteins such as
thrombin and fibrin.

H. Exotic proteins:
-Monellin is a protein that has a very
sweet taste; it is found in a certain
African plant.
-Resilin is an elastic protein in the
hinges of insect wings.

-There are 20 known amino acids and

can be arrange in almost infinite
number of sequences to make almost
an infinite number of different
proteins.
Structure
-Proteins can be considered as
polymers of amino acids. Amino acids
are linked by a covalent peptide bond
into linear chain, which is called
peptide or polypeptide chain.
When a polypeptide chain undergoes
folding, a protein is being formed. In
regard to complexity of molecule,
proteins fall in to simple and
conjugated.

-Mussels secrete glue proteins to

attach themselves to rocks.

PROTEINS

SIMPLE
MOLECULAR MASSES

-Amino acids are nitrogen-containing

organic compounds which form a
building blocks of proteins. They are
joined to each other by peptide
linkage; that is , the amino acids is
linked to the carboxyl group of another
amino acids by the removal water.
Thus, two amino acids form a
dipeptide, three amino acids forms a
tripeptide, and so on.
-Amino acids are any of a group of
organic molecules that consist of a
basic amino group (-NH2), an acidic
carboxyl group (-COOH), and an
organic R group (or side chain),
which is unique to each amino acid.
Although more than 100 amino acids
occur naturally, only 20 are commonly
used in protein synthesis. General
structure of the amino acids found in
proteins:

-The properties common to all amino

acids are due to the relative special
arrangements of the carboxyl and
amino groups. The physical and
chemical properties unique to each
amino acid are the result of the
structure and chemical properties of
the R group. Amino acids are
generally grouped according to the
polarity (the tendency to interact with
water at a neutral pH) and charge of
the R group
Amino Acids

General formula:
General formula:
H
R

COOH

NH2

R- hydrogen, aliphatic, aromatic or

heterocyclic components

A peptide bond is formed by a

condensation (water-loss) reaction
between the carboxyl group of one
amino acid and the amino group of the
next amino acid occurring in a protein.
Conceptually, protein structure can be
considered at four levels. The
primary structure consists of a
sequence of amino acids linked
together by covalent peptide bonds
and includes any disulfide bonds.
In humans, 8 amino acids cannot be
synthesized. They are considered as
essential amino acids including
isoleucine, leucine, lysine, methionine,
phenylalanine, threonine, tryptophan,
and valine, making them a

requirement in the diet. The remaining

are nonessential amino acids.
Amino acids are joined covalently by
peptide bonds to yield proteins
(including enzymes and hormones;
structural, transport, and contractile
elements; and molecules of special
biological activity).

Protein + lipid: lipoprotein.

Secondary structure refers to

regular, recurring arrangements in
space of adjacent amino acids in
polypeptide chain. There are few
common types of secondary structure,
the most prominent being the helix
and the conformation. Tertiary
structure refers to the spatial
relationship among all amino acids in
a polypeptide; it is the complete threedimensional structure of the
polypeptide. Proteins with several
polypeptide chains have one more
level of structure.

Derived Proteins

Classification
Proteins may be classified in number
of ways including physical and
chemical properties, physical shape,
and nutritional properties.

Simple and Conjugated Proteins

1) Simple proteins are composed
only of amino acids.
2) Conjugated proteins: proteins
which form complexes with other
substances
Protein + carbohydrate: glycoprotein.
Protein + nucleic acid: nucleoprotein.

Protein + a metal: metalloprotein.

Protein with a phosphate group
attached to one residues:
phosphoprotein.

Are substances resulting from the

decomposition of simple and
conjugated proteins by the actions of
heat and other physical forces or
hydrolysis agents. These includes the
rearrangement of within the
molecules without breaking the
peptide bond, such as that occurring
coagulation, and also substances
formed by hydrolysis of the protein to
smaller.
Ex: proteoses, peptones, and
peptides.
Physical Shape
Fibrous Proteins- consist of long
polypeptide chains bound together in
more or less parallel fashion to form a
linear shape. The four major fibrous
proteins:
a. Collagen ( most abundant protein
found in
vertebrates)
b. keratin
c. Fibrinogen
d. Muscle protein (actin and
myosin)

Classification
Based on the polarity of R and
interaction with water
1)Non-polar, aliphatic R group

2)Aromatic R group nonplar,

hydrophobic interaction , absorbs UV
Globular proteins are chains of
amino acids that are coiled and tightly
packed together in a round or
ellipsoidal shape. They are generally
soluble in fluids.
a. Insulin
b. Hemoglobin
c. albumin
d. globulin

3) polar, uncharged R group soluble

in water

ENERGY SOURCE
-If the diet does not provide enough
energy, the body must begin to break
down its own protein
-The proteins are broken down into
individual amino acids, then
deaminated, and the remaining
carbon, hydrogen, and oxygen
compounds are used to make energy
or glucose
-If the diet contains too much protein,
the excess will be converted to
glucose, or stored as fat

4) Positively charged (basic) R group

Primary structure number and

sequence of amino acid polypeptide
chain
By convention, it begins with a.a.
whose amino group is free ( N-terminal
end) and ends with carboxyl group (Cterminal end)
5) Negatively charged (acidic) R group

Secondary structure
-Shape (conformation) of molecule
-a-helix coiled spring
-maintained by intramolecular
bonds

H-

-b-pleated sheath maintained by

intermolecular H-bonds

Tertiary structure
-Specific folding and bending - side
chain is responsible (R group)

4 categories of protein structure

-H-bonding, Salt bridges, disulfide

bridge, hydrophobic interaction
responsible for stability

-2 major groups : fibrous and globular

8. Tryptophan
9. Valine
Nutritional Properties
Essential amino acids are those
that cannot be synthesized in the body
at a rate sufficient to meet the growth
and maintenance requirements. They
must provided preformed in the diet.

Quaternary structure

Nonessential amino acids- are

those that can be synthesized in the
body from an available source of
nitrogen and a carbon skeleton. They
mixed diets contain ample amount of
both essential and nonessential amino
acids.
Base on their amino acids content ,
foods are often classified as sources
of:

Nonessential Amino Acids

1. Alanine
2. Arginine*
3. Aspartic acid
4. Cysteine*
5. Cystine
6. Glutamic acid
7. Glutamine*
8. Glycine
9. Proline
10. Serine
11. Tyrosine
Classification of Amino Acid
Essential Amino Acids
1. Histidine
2. Isoleucine
3. Leucine
4. Lysine
5. Methionine
6. Phenylalanine
7. Threonine

Complete protein - contains all the

essential amino acids in proportions
capable of maintaining life and
supporting a normal rate of growth.
(high biologic value proteins) ex. Egg,
milk, meat , fish and poultry.
Partially complete proteincontains all the essential amino acids
but a relatively small amount of some
of the amino acids necessary for
growth. Ex. Gliadin and hordein.
Totally incomplete protein lacks
1 or more of the essential amino acids
therefore incapable of replacing or
building new tissues, hence cannot
support life or growth. Ex. Are zein or
gelatin.
Amino acids have a great variety of
chemical reactive groups providing a
wide range of reactivity proteins. The

reactions for individual side-chain

radicals and -amino and -carboxyl
groups are specific both for free amino
acids and proteins, whereas the
reaction for peptide group is
characteristic of the proteins and
peptides. Specific reactions are used
for the purpose of identifying amino
acids and proteins in biological media,
for qualitative and quantitative
analysis.
Biuret Reaction
The biuret reaction is a method that
can be used to determine the amount
of soluble protein in a solution. The
biuret reagent (coppersulfate in a
strong base) reacts with peptide bonds
(at least two peptidebonds) and
changes color when it does so. The
spectrophotometer can then be used
to measure the intensity of the color
produced. The more protein present
the darker the color.
Ninhydrin and Xanthoprotein Reaction
Ninhydrin reaction
The reaction is typical for -amino
acids. They react with ninhydrin
reagent yielding CO2, aldehyde and a
blue colored complex compound:
Xanthoprotein reaction
This reaction is specific for cyclic
amino acids such as phenylalanine,
tyrosine, tryptophane and histidine.
Aromatic amino acids react with HNO3
yielding a yellow nitrocompound,
which changes color to orange in
alkaline medium owing to the
formation of salt.

Millons Reaction
It is used for the determination of
tyrosine content in proteins. Millon
reagent is composed of salts of
mercury dissolved in HNO3. Tyrosine
(both free and constitutive of proteins)
reacts with reagent and produces a
purple-red salt of mercury:

Precipitation of Proteins
A number of proteins, including most
globulins, are water-soluble. The
hydration of molecules makes the
solutions of these proteins stable. The
solubility of protein solution may be
disturbed by factors inducing
precipitation. The precipitation of
protein may be reversible and
irreversible.
Precipitation is considered as
reversible if native molecular
conformation is preserved. In this case
the functional activity of protein can
be recovered after dissolving the
precipitate. Reversible precipitation is
induced by acetone, ethanol, and a
high concentration of mineral salts.
Chemical modification of side chains
of amino acids or splitting of peptide
bonds always induces an irreversible
precipitation called denaturation.
Proteins may be denatured by heating,
high-concentrated urea, detergents,
extreme pH, many organic solvents,
and radiation. Denatured protein loses
its native conformation and biological
activity.
Precipitation by Heating
When proteins are exposed to

increasing temperature, loss of

solubility or enzymatic activity occurs
over a fairly narrow range. Depending
upon the protein studied and the
severity of the heating, these changes
may or may not be reversible. Few
proteins can remain biologically active
above 50oC. As the temperature is
increased, a number of bonds in the
protein molecule are weakened.
Precipitation by salts of heavy
metals
Heavy metal salts usually contain
Cu2+, Hg+2, Pb+2, Ag+1 Tl+1, Cd+2 and
other metals with high atomic weights.
Since salts are ionic they disrupt salt
bridges in proteins. The reaction of a
heavy metal salt with a protein usually
leads to an insoluble metal protein
salt. This reaction is used for its
disinfectant properties in external
applications. For example AgNO3 is
used to prevent gonorrhea infections
in the eyes of newborn infants
Precipitation by Tannins and Picric
Acid
Both reagents react with heterocyclic
amino acids of proteins and produce a
precipitate.
Chromatography of amino acids
Partition paper chromatography is
widely employed for the determination
and separation of amino acids. The
solvent migrates along a strip of paper
and carries amino acids dissolved in it.
The distance of run of different amino
acids depends on the constant termed
the distribution coefficient (Rf). This
coefficient is the ratio of migration

distance of given amino acid (A) to the

migration distance of solvent front
Proteins make up 68% of the blood.
Amongst them, are proteins that
leak from dead and dying cells,
proteins secreted into blood or
released from tumors. Identifying
these proteins allows scientists to
conduct additional studies to define
functional role of each protein in cells
and the body. After blood is withdrawn
from a vein and allowed to clot, the
clot slowly shrinks. As it does so, a
clear fluid called serum is squeezed
out. Serum is blood plasma without
fibrinogen and other clotting factors.
Blood serum contains two major
protein groups - albumin and
globulin, which are separated by
salting-out technique. Both albumin
and globulin carry substances through
the bloodstream. Using protein
electrophoresis (separation of protein
molecules according to their charge),
these two groups can be separated
into five smaller groups (fractions):
Albumin. It is a major group of
related proteins produced in the liver.
In addition to carrying substances
through the bloodstream, albumin
proteins help to keep water from
leaking out of tiny blood vessels
(capillaries). Albumin may also help
with tissue growth and healing. More
than a half of the protein in blood
serum is albumin.
Alpha-1 globulin. High-density
lipoprotein (HDL), the "good" type of
cholesterol, is included in this fraction.
Alpha-2 globulin. A protein called
haptoglobin binds to oxyhemoglobin

yielding a high molecular mass

complex, which cannot be filtered by
the kidneys. In regard to this,
haptoglobin can be considered as a
natural antibody against hemoglobin.

contains all the essential amino acids,

as long as all the essential amino
acids are eventually present in the
diet: see "Complete protein" and
"Protein combining".

Separation of Proteins: gel- filtration

Factors Affecting Protein Requirements

Molecular sieve column

chromatography is used to separate
proteins or their fragments. This
method is based on the differences in
molecular weight and shape of
proteins. Small and large molecules of
proteins are separated on the column,
because small molecules move in and
out of the porous particles of the
matrix (they are retained by the gel),
meanwhile large molecules move only
outside the matrix particles and
therefore run down a column with the
solvent. High-hydrated polymers of
polysacharides serve as the matrix.
They are available in different sizes for
separation of proteins of different
molecular weights.
In nutrition proteins are broken
down in the stomach during digestion
by enzymes known as proteases into
smaller polypeptides to provide amino
acids for the body, including the
essential amino acids that the
organism cannot be biosynthesized by
the body itself. Thus, protein from
one's diet should provide both
essential and non-essential amino
acids for protein synthesis.
Most animal sources and certain
vegetable sources have the complete
complement of all the essential amino
acids in adequate proportions.
However, it is not necessary to
consume a single food source that

1. Body weight
2. Growth
3. Pregnancy
4. Lactation
5. Aging
6. Nutritional status
7. Protein quality
Sources of Protein
Animal Sources:
- eggs, milk, and milk products,
meat , fish, poultry, and sea foods.
Plant Sources:
- cereals( wheat, rice ,corn);
legumes(mongo beans, peas, soy
beans, white beans, red beans, lentils,
lima beans, nuts, processed
vegetables proteins like tofu.

Protein Malnutrition
Kwashiorkor ( meaning the
displaced child) occurs in children
shortly after weaning when the diet is
high in starch and low in quality and
quantity of protein.
Marasmus ( wasting or
withering). Is usually seen at a

somewhat earlier age than

kwashoirkor.