Chapter 3

THE BIOLOGICAL MOLECULES

Module 5: AMINO ACIDS, PROTEINS and NUCLEIC ACIDS

Lesson 1
Course Intended Learning Outcomes
At the end of the chapter, the student should be able to:
1. Identify the elements that make up the amino acids, proteins and nucleic acids;
2. Describe the chemical composition and functions of amino acids, proteins and nucleic acids;
3. Differentiate the different classes of amino acids, proteins and nucleic acids;
4. Differentiate RNA from DNA;
5. Determine the recommended daily allowance of proteins;
6. Determine the effect of insufficient or oversupply of proteins in the body;
7. Determine the common food sources of proteins; and
8. Describe the formation and breakdown of amino acids, proteins and nucleic acids in the body.

Definition
Proteins are compounds of high molecular weight consisting largely of chains of amino acids united in
peptide linkage. It is a component of living cells and is synthesized by the cell to effect growth. It is composed
of about 50% carbon, 7% hydrogen 23 % oxygen, 16% nitrogen, 0 – 3 % sulfur, 0 – 3% phosphorus.

Upon hydrolysis, protein will be broken into its basic building blocks, the amino acids.

Amino Acids

The amino acids are the building blocks of proteins. They are characterized by an amino group in an
alpha position to the carboxyl group.

The Structure of the Amino Acids

NH3 + where:
l R = hydrogen, an aliphatic, aromatic or heterocyclic radical.
H – C – COO –
l
R

The alpha carbon of the amino acid has four different groups attached to it. As a result, the carbon atom
is assymetric or a chiral carbon., and therefore makes the amino acid optically active. Of the 20 amino acids, 19
are optically active.
Glycine is a symmetrical amino acid because its alpha carbon is attached to two hydrogen atoms. It is
therefore not optically active.

Classification of Amino Acids:

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I. Classification based upon its ability to interact with water.

1. Neutral non-polar amino acids – contain hydrocarbon R – groups. These amino acids are referred to
as neutral because its R – groups do not carry any charges. They interact poorly with water.
(hydrophobic).
These are:

Glycine Phenylalanine
Alanine Tryptophan
Valine Methionine
Leucine Cysteine
Isoleucine Proline

2. Neutral polar amino acids – contain functional groups capable of forming hydrogen bonds and acn
therefore interact with water (hydrophilic).

Serine Asparagine
Threonine Glutamine
Tyrosine

3. Acidic amino acids – contain the carboxylate (carboxylic acid group) side chains such as aspartic
acid and glutamic acid.

4. Basic amino acids – carry positive charge at physiological pH. They form ionic bonds with acidic
amino acids.
Lysine
Arginine
Histidine

II. Classification according to R – group.

1. Aliphatic amino acids
Glycine
Alanine
Leucine
Isoleucine
Valine

2. Aromatic amino acids

Phenylalanine
Tyrosine
Tryptophan

3. Hydroxyamine acids
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 Serine
Threonine

4. Acidic amino acids

Aspartic acid
Glutamic acid

5. Basic amino acids

Lysine
Arginine
Histidine

6. Sulfur amino acids

Cystine
Cysteine
Methionine

7. Imino Acids
Proline
Hydroxyproline

III. Classification according to essentiality

1. Essential amino acids - are those amino acids which cannot be synthesized fast enough for the
needs of the body and must be supplied therefore in the dietary proteins.

Arginine Leucine Phenylalanine

Histidine Lysine Threonine
Isoleucine Methionine Tryptophan
Valine

2. Non – essential amino acids – are those amino acids which can be synthesized biologically from
other nutrients.

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4
PHYSICAL PROPERTIES OF AMINO ACIDS
Amino acids appear as white crystalline solids soluble in cold water except cystine and tyrosine
which are soluble in hot water. They are insoluble in alcohol except proline and hydroxyproline. Most of
them are sweet. Leucine is tasteless while arginine is bitter.

CHEMICAL PROPERTIES OF AMINO ACIDS

1. Amino acids are charged particles

They are therefore not true amino acids but rather ionized internal ammonium salts. This form of an
amino acid is called ‘Zwitterion.” Fundamentally, acids react with the acidic groups of amino acids and
bases react with the basic group. Amino acids therefore are classified as amphoteirc since they can act
with both acid and base.

When the amino acid is in the “zwitterion” state, it will not migrate to either electrode in an electric
field. The pH at which the amino acid has no tendency to move to either positive or negative electrodes
is called the isoelectric point.

When an acid is added to an amino acid, the latter assumes its positively charged form and moves
toward the cathode, and when a base is added to an amino acid, the latter assumes its negatively charged
form and moves toward the anode. This migration is called electrophoresis.
2. Amino acids form esters with alcohol and hydrochloric acid. This property was made use by Fischer,
who separated amino acids from each other by fractionally distilling their esters.
3. Amino acids can be acetylated, benzylated or methylated. These reactions are used in the body for
detoxifying purposes.
4. All amino acids, except proline and hydroxyproline react with nitrous acid with the liberation of
nitrogen gas. This is involved in the Van Slyke method for the determination of amino groups in
proteins, blood and other biological materials.

5. Formol Titration (Sorensen Reaction) – free amino acids and proteins can be estimated by a technique
known as formol titration. The reaction occurs between amino groups and formaldehyde. This is made
use of in following the course of protein digestion in which amino acids are liberated.

6. Decarboxylation causes the formation of primary amines.

H H
I I
R – C – COOH - CO2 R–C–H + CO2
I I
NH2 NH2

COLOR REACTIONS OF AMINO ACIDS

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General Reactions:

1. Ninhydrin Reaction – when alpha amino acids are treated with “ninhydrin” (triketohydrindene
hydrate), in aqueous solution, color is developed ranging from clear, deep blue to a violet –pink or
even red. Ninhydrin is probably the most delicate reagent for detecting the presence of protein or of
an aplha-amino acid (except proline and hydroxyproline).

2. Folin’s Reagent – in the presence of strong alkalies and of amino acid, beta – naphthoquinone
sulfonic acid develops a striking, deep red color. However, urea, uric acid, creatinine and hippuric
acid do not yield colors.

Specific Reactions:

1. Millon Reaction – a red color develops when an alpha amino acid, a protein or a protein hydrolysate
is heated with a solution of mercurous and mercuric nitrate or nitrite. The red coloration is specific
for the phenol group. The reaction is specific for tyrosine.

2. Xanthoproteic Reaction – when an alpha amino acid is treated with nitric acid, a yellow coloration is
produced which is intensified to orange on addition of ammonium hydroxide. The yellow color so
commonly produced on skin which has come in contact with nitric acid is an example of the
xanthoproteic reaction and indicates the presence of nitrated protein. Nitration takes place in the
benzene ring, and the reaction is, therefore, specific for aromatic nuclei which are easily nitrated
tryptophan, tyrosine or phenylalanine.

3. Tryptophan Reaction – when tryptophan or protein is heated with strong sulfuric or hydrochloric
acid in the presence of aldehyde, a blue or violet color is produced. The color is due to the
condensation of the alpha – hydrogen of the indole nucleus with an aldehyde.

The test is known under a variety of names depending on the aldehyde employed:

a. Liebermann reaction – many proteins give the test without the addition of an aldehyde, owing
either to
the contamination or to the presence of carbohydrate units in the protein molecule.

b. Acree – Rosenheim – when formaldehyde is added to the reaction.

c. Adamkiewicz reaction – glacial acetic acid is added because of glyoxalic acid content, COOH –
CHO.

d. Ehrlich reaction – p – dimethylaminobenzaldehyde is used.

e. Cole’s test – the test uses the unsubstituted benzaldehyde

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 f. Hopkins – Cole test – uses preformed glyoxalic acid.

4. Reduced Sulfur Test – a black coloration is produced when a protein or a mixture of amino acids is
heated with alkali and lead acetate. The coloration is due to the formation of black lead sulfide and
appears to be specific for the presence of cystine or cysteine, or rather the groups – S – S – or SH
radical.

5. Sullivan Cystine Reaction – the test consists essentially in adding to the cystine hydrochloride 1 – 2
ml of a 1% sodium cyanide solution made up in 0.8 N NaOH, then adding 1 ml of a fresh 0.5%
aqueous solution of 1,2 – naphthoquinong – 4 – sodium sulfonate, then adding 5 ml of a 10 – 20%
Na2CO3 solution made up in 0.5 N NaOH, and allowing the mixture to stand at 20oC for 30 minutes.
Pure red color is then developed by adding 1 ml of a 2% aqueous solution of sodium hyposulfite.
The color is specific for cystine or cysteine.

6. Sakaguchi Test – this test consists in adding 5 ml of a 1% solution of a protein, or a 1% solution of

an amino acid mixture to 2 ml of a 15% sodium hydroxide followed by a 5 ml of a 0.15% solution of
alpha-naphthol. The entire mixture is treated with 0.3 N sodium hypochorite. After standing at 2oC
to 4oC for 40 minutes, an intense red color is developed. Only compounds containing the free
guanidine group react. Accordingly, the color test is specific for arginine.

7. Diacetyl Test – if a dilute solution of protein or amino acid is mixed with small amount of 10%
KOH solution and then one drop of a 1% solution of diacetyl (CH3 – CO – CO – CH3) is added, a
pink color with a green fluorescence is developed. This reaction is obtained in the presence of
arginine.

8. Nitroprusside Reaction – proteins containing cysteine give a reddish color with sodium nitroprusside
in dilute ammoniacal solution. This reaction is due to the presence of SH group.

9. Bromine – Water Reaction – tryptophan gives pink color when treated with bromine water.

THE FUNCTIONS OF PROTEINS

Among all the biomolecules, proteins have the variety of functions. These are:

1. Structure, protection and support. Examples of which are collagen (connective tissues), fibroin (silk
protein), elastin (blood vessels and skin).

2. Defense and protection against chemical and mechanical injury. Keratin in the skin provide
protection. Fibrinogen and thrombin induce blood clotting. Antibodies of immuno-globulin protects
man from infection.

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 3. Regulation through the body hormones. Examples of which are insulin, glucogen, growth and sex
hormones to name a few.

4. Catalysis through enzymes. Enzymes are proteins that direct and regulate the speed of biochemical
reactions. Examples are amylase, oxidase, phosphatase, etc.

5. Transport as carrier of ions or molecules across cell membranes or between cells. Other transport
proteins are hemoglobin, carrying oxygen to the tissues from the lungs; and lipoproteins, LDL and
HDL, that transport lipids from the liver and intestines to other organs.

CHEMICAL STRUCTURES OF PROTEINS

The building units of protein molecules are alpha – amino acids of the L – series. These amino
acids are joined together by bonds between the carboxyl group of one amino acid and the amino group
of another, with splitting off of water;

H O H H H O H H
I II I I I II I I
R – C – C – OH + H–N–C–R R – C – C ---------------- N – C – R
I I I I
NH2 COOH NH2 COOH

Peptide Linkage

The bond between two amino acids, - CO – NH – is called peptide bond or amide bond. When just two
amino acids are involved the resulting compound is called a dipeptide. The availability of an amino and
carboxyl group in the dipeptide makes further combinations with other (or the same) amino acids possible, so
polypeptides maybe built.

Other bonds also play important roles in the protein molecule. Intramolecular bonds are formed when
the disulfide group of cystine from cross linkages resulting in ring systems within the protein molecule. A salt
bridge or ionic bond is formed between groups which are positively and negatively charged. Hydrophobic
bonds are formed by amino acids like leucine, valine, phenylalanine, tryptophan and proline which adhere to
each other forming a “micelle” and which do not mix well with other. Hydrogen bonds result from the attraction
of electronegative atoms in the protein molecule.

DIFFERENT STRUCTURES OF PROTEIN

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Primary Structure of Protein

Several levels of structure of the protein molecule have been recognized. The first level of protein
structure, the primary structure, is formed when acid molecules are linked together through the covalent peptide
bonds. These bonds are formed between the carboxyl group of one amino acid with the amino group of
another. Three or more amino acids linked together forms a polypeptide chains.

Secondary Structure of Protein

Hydrogen atoms in the polypeptide chain can bond with oxygen and other atoms of different amino
acids. The H – bond causes the chain to twist into a coil or a helix or a pleated – like chain of amino acids

Ternary Structure of Protein

The different R – groups of the amino acids in the polypeptide chain may interact causing the chain to
twist, bend and fold into a three-dimentional shape. This results into the tertiary protein structure which more
specifically maybe formed and stabilized by:
a. Disulfide bonds of cysteine
b. Hydrogen bonds formed by the phenol group of tyrosine with the carboxyl group of aspartic and
glutamic acid.
c. Salt bonds formed by ionizable functional groups of amino acids.
d. Proline and hydroxyproline which are secondary amines and not primary amines like the other amino
acids. They do not have a peptide bond hydrogen to contribute to hydrogen bonds. This cause a break in
the polypeptide chain.
e. Van der Waals forces refer to weak hydrophobic electrostatic interaction between dipoles of R – groups
in proteins.

Quaternary Structure of Protein

Some proteins are able to incorporate its polypeptide units through disulfide linkages or hydrogen
bonding and Van der Waals forces. The resulting protein can be fibrous or globular or a combination of both.
An example of this is hemoglobin.

CLASSIFICATION OF PROTEINS

I. According to Gross Structure

Simple Proteins
A. Fibrous Proteins or Scleroproteins (albuminoids, sclerins)
⮚ Largely insoluble in ordinary aqueousmedia (salt solutions, acids, etc)
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 ⮚ Molecular weights are high
⮚ Consists of fibers made up of long linear molecules arranged (roughly) paralleled to the fiber axis.
⮚ Function is for structure and support
⮚ Examples are collagen from from cartilage; myosin from muscle; keratin from hair; fibrin from
blood clot.

B. Globular Proteins
⮚ Soluble in aqueous media (salt solutions, acids , bases, acids, aqueous alcohol)have been
crystallized and have definite molecular weights
⮚ Can be denatured
⮚ Examples are enzymes and hormones

II. According to Solubilities

A. Albumins
⮚ Soluble in water
⮚ Coagulated (change to solid) on heating
⮚ Examples are egg albumin; serum albumin; lactalbumin from milk; leucosin from wheat

B. Globulins
⮚ Insoluble in water; soluble in dilute salt solutions but precipitated when the salt concentration is
increased. Salt used: NaCl, MgSO4, (NH4)2SO4
⮚ Coagulated on cooking
⮚ Examples are myosinogen from muscles; edestin from hemp; ovoglobulin from egg yolk; serum
globulin, amandin from almonds; legumin from peas; exelsin from Brazil nuts.

C. Glutelins
⮚ Insoluble in neutral solvents but soluble in dilute acids and alkalis
⮚ Examples are glutelins from wheat; oryzenin from rice

D. Alcohol – Soluble Proteins (Prolamins and Gliadins)

⮚ Soluble in 70 – 80% alcohol; insoluble in water and absolute alcohol
⮚ Examples are gliadin from wheat, hordein from barley; zein from corn

E. Fibrous Proteins
⮚ See above IA

F. Histones
⮚ Soluble in water; insoluble in ammonia
⮚ Preicipitated by solutions of other proteins
⮚ Goagulum formed on heatin is souble in dilute acids
⮚ Examples are globulin from hemoglobin

G. Protoamines
⮚ Polypeptides which are less complex
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 ⮚ Soluble in water
⮚ Not coagulated by heating
⮚ Precipitates other proteins from their aqueous solutions
⮚ Possess strong basic properties
⮚ Form stable salts with strong mineral acids
⮚ On hydrolysis, forms amino acids with basic character
⮚ Examples are salmine from salmon; sturine from sturgeon; clupeine from herring; scrombine from
mackerel; cyprinine from carp

III. According to Content of Non-Protein Constituents

A. All of the Proteins in II

B. Conjugated Proteins – substances made up of proteins combined with some other compound or
compounds.

1. Nucleoproteins
⮚ combination of proteins with nucleic acids
⮚ examples are products obtained from glandular tissue and from the germ of grain.

2. Glycoproteins
⮚ combination of protein with carbohydrates (such as hexoses, hexosamines and hexuronic acids)
⮚ examples are mucin from saliva; osseomucoid from bone; tendomucoid from tendon

3. Phosphoproteins
⮚ Combination of protein with phosphorus containing substances other than nucleic acid or lecithin
⮚ Examples are casein from milk; vitellin from egg yolk

4. Chromoproteins
⮚ Combination of protein with various pigments
⮚ Examples are hemoglobin from blood pigment; ferritin from the liver and spleen; catalase,
peroxidase and cytochrome C (iron-containing enzymes); hemocyanin (copper-protein complex)

5. Lipoproteins
⮚ Combination of proteins with lipids
⮚ Occurs in cell nuclei, blood, egg yolk, milk, serum, etc.

IV. According to State of Degradation

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A. Native Proteins
⮚ Proteins in the state in which they exist is the intact cell

B. Derived Proteins

⮚ Products of protein degradation, the first step of which is denaturation. These are:
1. Primary Protein Derivatives – products of comparatively slight hydrolytic change in the protein
molecule:

a. Proteans – insoluble products resulting probably from the action ( for a comparatively short
time) of hot water, dilute acids or enzymes. Examples are myosan from myosin; odestan
from edestin
b. Metaproteins (infraproteicns) – products of further action of acids and alkalies and are
insoluble in solutions of neutral salts, but soluble in dilute acids. Examples are acid
metaprotein or acid albuminate and alkali metaprotein or alkali albuminate.
c. Coagulated proteins – insoluble products resulting either from the action of heat or alcohol.

2. Secondary Protein Derivatives – products of a more extensive hydrolysis of the protein. These
are:

a. Proteoses – soluble in water; cannot be coagulated on heating; precipitated by saturating their

solutions with (NH4)2SO4.
b. Peptones – soluble in water; not coagulated on heating; not precipitated by saturating their
solutions with (NH4)2SO4; not precipitated by certain alkaloidal reagents such as
phosphotungstic acid.
c. Peptides – combination of two or more amino acids, the carboxyl group of one amino acid
being joined to the amino group of another.

V. Accoording to Function

A. Structural - myosin from muscle

B. Storage - ferritin in the liver stores iron
C. Enzymes - pepsin of the stomach
D. Hormones - insulin, vasopressin
E. Antibodies
F. Toxins
G. Special purpose - hemoglobin to carry molecular oxygen; rhodopsin in the visual process

VI. According to Nutritional Completeness

A. Complete – maintaining life and providing for normal growth when used as a sole protein food.
Examples are casein and lactalbumin from milk; ovalbumin and ovovitellin from egg; glycinin from
soybean; excelsin from Brazil nut; edestin, glutenin and maize glutelin of grains.

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 B. Partially Incomplete - maintaining life but not supporting normal growth. Examples are gliadin from
wheat; herdein from barley; prolamin from rye.
C. Incomplete – incapable of maintaining life or supporting growth when fed as the sole protein. Examples
are zein from corn; gelatin.

CHEMICAL PROPERTIES OF PROTEINS

1. A protein has a constant solubility regardless of the amount of substance present in the solid phase. This
property is a sensitive test for the presence of protein impurity.

2. Proteins are hydrolyzed by:

a. Acids – 20% HCl, 25% H2SO4, dodecylsulfonic acid, picric acid. The organic acids are more
effective than the mineral acids in liberating the NH3 from its amides. Hydrolysis is proportional to
the nydrogen ion activity of the acid solutions.
b. Alkalies – NaOH, KOH, and Ba(OH)2 cause very rapid and complete hydrolysis but they bring about
decomposition, especially the deamination of certain amino acids. Unfortunately, alkaline hydrolysis
result in the racemization of the amino acids so that they are isolated to a large extent in the optically
inactive form.
c. Enzymes – pepsin hydrolyzes proteins to proteoses, peptones and polypeptides only. Trypsin and
and ereptic enzymes hydrolyze protein to amino acids but the process is slow and incomplete.

3. Proteins are precipitated by:

A. Acids
Proteins react with acids because of their free amino groups. The acids which precipitate the proteins
in this way are sometimes termed “alkaloidal reagents” because they precipitate many of the
alkaloids. Some innorganic acid precipitants are HNO 3 and HClO4. Some organic precipitants are
trichloroacetic acid, molybdic, phosphotungstic, picric and tannic acids.

Picric acid is used in treatment of burns because it produces an astringent effect on the tissue,
diminishes secretion of the mucous membranes and prevents absorption of toxins.

Tannic acid is used to relieve diarrhea. Commercially, it is used to precipitate collagen in hides, thus
yielding leather. It is also incorporated in an ointment for burns, producing a protective crust of
protein tannate over the delicate area.

Sulfosalicylic acid is used to detect proteins quantitaively and determine them in the urine and body
fluids.

1. Heller’s Test
Reagent: HNO3
Result: white precipitate turning to yellow

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 2. Robert’s Test:
Reagent: HNO3, MgSO4
Result: white precipitate

B. Salts of Heavy Metals

Precipitation occurs on the alkaline side of the isoelectric point. The metals unite with the carboxyl
group, thus forming proteinates (e.g.,Hg-, Ag-, Pb-, Cuproteinates).,

AgNO3 is used in cauteries. It precipitates the proteins of tissues as silver salts.

Antidotes for metallic poisoning: egg white, milk and other liquid protein. The metallic precipitate
must be removed from the stomach by an emetic or by a stomach tube to prevent the digestion of the
protein and the liberation, re-solution and absorption of the poisonous metal.

C. Neutral Salts
(NH4)2SO4, Na2SO4, and NaCl are used to precipitate or “salt out” proteins. Precipitation maybe due
to dehydration of molecular aggregates in solutions.
D. Alcohol
The addition of alcohol to protein in solution has two effects depending on the concentration of the
solution:
1. If the solution is dilute (aqueous), the alcohol reduces the solubility of the proteins due to an
increase in the electrical forces between charged particles in solution.
2. At high concentration, the protein molecules are dehydrated thus causing precipitation.
4. Proteins form complexes with:
A. Nucleic acids to form nucleoproteins
B. Carbohydrates to form glycoproteins
C. Phosphorous – containing substances other than nucleic acid or lecithin to form phosphoproteins.
D. Pigments to form chromoproteins
E. Lipids to form lipoproteins
F. Enzymes and coenzymes

5. Proteins unite with certain dyestuffs. Among them are picric acid and flavianic acid.
6. When burned, proteins decompose and liberate a characteristic odor of burned hair or feather.
It blackens to prove the presence of carbon.

COLOR REACTIONS OF PROTEINS

Because proteins are composed of alpha amino acids, they exhibit the same color reactions as the amino acids.
A specific color test for proteins is the BIURET test:

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 Reagents: NaOH, CuSO4
Result: violet color
Indication: presence of a peptide linkage or other closely related group such as

-C –N or the –C–N–
I I II I
S H NH H

Free amino acids will not give the Biuret test. Ammonium salts interfere with this test by forming a deep blue
compound. A single peptide linkage will not give the Biuret reaction. The color is due to a complex in which
copper is coordinated with four hydrogen atoms, therefore at least 2 peptide linkages is required to produce a
positive result.

PROTEIN DENATURATION

Denaturation is the change in the chemical, physical and biological properties of a protein from those of
the native state, characterized by an unfolding of the molecule from its previous configuration.

Kinds of Denaturation:

1. Unfolding of the peptide chains, e.g., action of urea on albumin

2. Dissociation of the protein into smaller units, which may or may not unfold, e.g., action of urea on the
tobacco mosaic virus molecule

Denaturing Agents:

1. Physical Agents

a. Heat: water is necessary for denaturation and coagulation by heat. The steps in heat coagulation are:
1st – denaturation; 2nd – flocculation; 3rd – coagulation.

b. Light: clouding of the lens of the eye in old-age cataract is probably due to denaturating of the
globulins present in the lens. Glass workers are particularly subject to cataract presumably as a result
of the infrared rays emanating from the molten glass. Change of fibrinogen to fibrin in blood cotting
may be partly due to light.

c. Surface tension
d. High pressure
e. Mechanical agitation

2. Chemical Agents

a. Organic solvents (alcohol, acetone, ether)

b. Acids and alkalies
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 c. Salts of heavy metals
d. Enzymes
e. Detergents
f. Urea and guanidine

ALTERATION OF PROPERTIES DUE TO DENATURATION

1. Physical
a. Increase in the viscosity of the solution
b. Increase in the rate of diffusion
c. Cannot be crystallized
d. Decrease in solubility

2. Chemical
a. Some groups in the denatured protein molecules become exposed and readily detectable. Among
these groups are the sulfhydryl (-SH), disulfide (- S – S - ) and the phenolic group. They are shielded
in the native state. Denaturation decreases the solubility of the proteins.

3. Biological
a. Increased digestibility by proteolytic enzyme has been found in certain denatured proteins.
b. Enzymatic or hormonal activity is usually destroyed.
c. Modification of the specific immunological properties.

DETECTION AND DETERMINATION OF PROTEINS

1. Heat Coagulation
Albumin is often detected in urine by the appearance of a coagulum on heating. The addition of acetic
acid to the heated sample will intensify the protein precipitate. Heat coagulation is sometimes used for
the gravimetric determination of proteins.

2. Precipitating Agents
In Heller’s Ring Test, protein in urine is detected by layering the urine over concentrated nitric acid and
observing the ring of acid-denatured protein which appears at the interface of the two liquids.

3. Specific Gravity
The specific gravity of plasma or serum is dependent on its protein content. A method for the
determination of plasma proteins involves the timing of the rate at which a drop of plasma falls through
a column of organic solvents of a density slightly less than that of plasma.

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BIOLOGICAL SIGNIFICANCE OF PROTEINS

Proteins have many functions. They include:

1. Providing the structural framework of tissues, membranes and cell walls.
2. Maintaining osmotic integrity.
3. Storage of some particular element, e.g., iron in ferritin
4. Enzymes, to catalyze biological reactions.
5. Hormones, to regulate metabolic processes, e.g., insulin
6. Carriage of molecular oxygen, e.g., hemoglobin
7. Transportation of lipids, e.g., lipoproteins
8. Taking part in antibody formation.
9. Acting to a certain extent as buffers in biological processes.
10. Secondary source of energy

Common Signs and Symptoms of a Protein Deficiency

While most people consume enough protein through their food choices on a day-to-day basis, those who
under eat, are vegetarian or vegan, or live in developing countries may not. The following are 13 signs of a
protein deficiency to be mindful of.

1. Muscle Wasting
When the body isn’t getting enough protein from food sources, “the body tends to take protein from
skeletal muscles to preserve more important tissues and body functions.” This condition is known as muscle
wasting, which can lead to a loss of energy and strength and may make doing physical activities more
challenging. Also, elderly people are especially susceptible, with “even moderate protein insufficiency” causing
muscle degeneration.

2. Bone Fractures or Breaks

In addition to your muscles, an inadequate amount of protein can also affect the health of your bones. In
fact, researchers stated that “not consuming enough protein may weaken your bones and increase the risk of
fractures.” This is because “Protein is needed for calcium absorption and helping with bone metabolism”.
Unfortunately, should you experience a fracture or break, a deficiency can slow recovery, as protein is essential
for healing, as well as building new cells, tissue, and skin.

3. Poor Sleep
Having trouble sleeping? It may be a sign that you’re deficient in protein. As the nutrient is crucial for
helping to keep blood sugar levels stable, not getting enough of it can result in swings that can “carry over
through the night,” resulting in poor sleep and insomnia. Additionally, protein “acts as the building blocks of
tryptophan, an amino acid that causes drowsiness.” Try eating protein-rich foods near bedtime to help stimulate
its production so you can get a better night’s sleep.

4. Brain Fog

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 Your body requires a certain amount of protein in order to keep your brain functioning properly. If you
find yourself experiencing brain fog, lack of motivation, poor concentration, or trouble learning new things, it
may be a sign that you’re deficient in the nutrient. Specifically, protein is essential to the production of
dopamine, epinephrine, norepinephrine, and serotonin, neurotransmitters that are needed to help you focus.
Researchers explain that these neurotransmitters “are synthesized in the brain using amino acids,” which are
created through when the body digests the nutrient.

5. Digestion Issues
Amino acids—which are created through the digestion of protein—are essential to a variety of digestive
system functions. Therefore, if you’re not consuming an adequate amount of protein, you may find yourself
experiencing stomach pain, gas, diarrhea, or sometimes constipation. This is because “Without enough protein,
enzyme production and muscle contractions in your GI tract will suffer.”

6. Moodiness and Anxiousness

As mentioned earlier, when protein is digested in the body it creates amino acids. And amino acids are
what comprise neurotransmitters like dopamine and serotonin, which control your mood. So if you’re not
getting an adequate amount of protein, the production of these neurotransmitters can suffer, resulting in mood
changes and increased anxiousness. Simply increasing your intake can help to synthesize more of these
hormones and help to “bring on positive feelings like calm, excitement and positivity.”

7. Sugar Cravings
If you’re low on protein, you may find yourself battling constant cravings for snacks that are high in
sugar. The reason for this is because protein helps to keep your blood sugar levels balanced, so if you’re not
getting enough “your glucose levels will be all over the place, encouraging you to reach for a quick fix like
candy,” according to Women’s Health Magazine.
The problem is that carbohydrates digest quite rapidly, so after consuming these sugary snacks you’ll
experience a spike in blood sugar, followed by a crash that causes you to reach for even more of them. Adding
more protein to your diet will help to slow down digestion to avoid this cyclical process.

8. Weight Gain
As mentioned previously, sugar cravings are a common sign that you’re deficient in protein. If you give
in to those cravings on a consistent basis, it can oftentimes lead to weight gain.
While high-protein food sources sometimes have more calories than carbohydrates, eating more of them can
help to increase satiety, preventing overeating. “They also help stabilize your blood sugar, allow you to retain
more muscle which burns more calories all day, and can reduce cravings.”

9. Edema
Insufficient protein intake can also lead to a condition called edema, where the skin becomes swollen
and puffy due to fluid retention. This is especially common in the body’s extremities, such as the feet and
ankles.
Citing Harvard Health Publications, Reader’s Digest explains that this occurs because “proteins help to hold salt
and water in blood vessels; without enough protein, these fluids can seep into surrounding tissues.” Edema is
often characterized by skin that looks stretched and shiny, and “retains a fingerprint after being pressed for a
moment.”
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10. Fatty Liver
Another common sign of protein deficiency is a build up of fat in the liver cells. If left untreated, this
accumulation can sometimes develop into fatty liver disease; a condition that can cause “inflammation, liver
scarring and potentially liver failure.” Although it’s not entirely clear why fatty liver disease occurs in those
who are protein deficient, the source says, “…studies suggest that an impaired synthesis of fat-transporting
proteins, known as lipoproteins, may contribute to the condition.”

11. Hair Loss

As hair is made up of 90 percent protein (specifically a type called keratin), if you’re not getting enough
of the nutrient it can cause hair to become thin or fall out. Research explains that this happens because “your
body stops using protein for non-essential things like hair growth in an effort to preserve its stores.” So if you
find that your hair has become dry, changed color or texture, or that you’re losing more of it than usual in the
shower or when running your hands through it, it may be a sign that you need to up your protein intake.

12. Skin and Nail Problems

As protein helps with nail formation, not getting enough of it can result in changes such as “white bands
or brownish spots on the nails.” You may also notice that your nails become weak, brittle, or develop ridges in
them. The source adds that a protein deficiency can also affect your skin, as “Protein enables cell regeneration,
produces new cells and replaces dead ones.” So if you’re insufficient in the nutrient, your skin may become dry,
flaky, cracked, and burn easily when exposed to sunlight.

13. Frequent Infections

Protein is essential to the functioning of the body’s immune system, as it “protects your body and
defends against foreign substances such as bacteria and viruses.” When you’re not consuming an adequate
amount of the nutrient, your white blood cell count decreases, weakening the immune system and making you
more susceptible to illness such as influenza and the common cold. These illnesses are often more severe in
those who are low on protein.

Assignment No. 1 Date of Submission: December 2, 2020

A. Enumerate and get a picture of the common food sources of Proteins.

B. Fill in the table below:

Age Group Recommended Daily Allowance (RDA)

Adult: Men

Adult: Women

Pregnant Woman

Lactating Mother

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 Female, 20 – 30 y/o

Male, 20 – 30 y/o

Female, 13 – 19 y/o

Male, 13 – 19 y/o

Children, 12 y/o and

below

Lesson 2

NUCLEOPROTEINS AND NUCLEIC ACIDS

A. Nucleoproteins
Chemical Composition
From the standpoint of protein chemistry, the nucleoproteins are a class of conjugaed proteins. They
are made up of nucleic acid (non – protein or prosthetic group) and protein (usually prolamine or histone). The
two components are attached or linked by primary (or covalent salt-like electrovalent ) bonds.

Occurence
They are found in all living cells. They occur in living organisms – from the simplest viruses, bacteria
and single – celled plants to the most complex and highly – developed tissues. They are most easily isolated
from yeast and from tissues with closely –packed cells and large nuclei such as the thymus gland. The other
organs rich in nucleoproteins are the spleen, pancreas, liver, intestine, mammary gland, brain and kidney. The
viruses are found to be entirely made up of nucleoproteins. The chromatin material of cells are largely
composed of nucleoproteins.
The term “nucleic” or “nucleo” is derived from the erroneous belief that these substances occur only in
the nuclei of cells.

Biological Importance
The nucleoproteins, more than any other constituents of living matter are concerned with vital cellular
organization and function. They considered to play an important role in the processes of cell division,
reproduction and transmission of hereditary factors. They are involved in protein synthesis of the different
tissues. Sub – components of nucleoproteins are associated in energy transfers in all living organisms either
directly or as co-factors to enzyme to promote reactions that provide energy.

B. Nucleic Acids
Chemistry and Structure
The nucleic acids are substances of high molecular weight. They maybe considered as polynucleotides
(meaning several nucleotides joined together) and each nucleotide (mono – nucleotide – specifically) is made
up of components: H3PO4 (phoporic acid), sugar (pentose) and nitrogenous base (purine or pyrimidine).

The sugar portion of nucleic acids is found to be pentose: ribose or deoxyribose

The purine bases that enter into the composition of nucleic acids are adenine and guanine.

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 The pyrimidine bases are cytosine, uracyl and thymine.

The simple structural representation of the mononucleotide and polynucleotide maybe as follows:

Mononucleotide:

Base Pentose Phosphate

Polynucleotide

Base 1
Pentose 3 5
Phosphate

Base 1
Pentose 3 5
Phosphate

Base 1
Pentose 3 5
Phosphate

The nitrogenous base (purine or pyrimidine) is linked to the terminal carbon atom (1) of the sugar and
the individual nucleotide through phospho – diester – groups connected at carbon (3) of one of the sugar
molecule to a terminal carbon (5) of the next.

Hydrolysis of nucleic acids maybe represented as follows:

Nucleic Acid
(Polynucleotides)

Mononucleotide

H3PO4 Nucleoside

Nitrogenous base Sugar

Purine – Adenine or Guanine Ribose or Deoxyribose
Pyrimidine – Cytosine, Uracyl
or Thymine

A nucleoside is a sugar-based compoud or a nucleotide minus H3PO4.

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Types of Nucleic Acids

There are two types of nucleic acids now definitely identified. Their names are derived from their sugar
component:
(1) Ribonucleic Acid (RNA); (2) Deoxyribonucleic Acid (DNA). The products of their hydrolysis are listed
below:

RNA DNA
1. H3PO4 1. H3PO4
2. Ribose 2. Deoxyribose
3. Adenine 3. Adenine
4. Guanine 4. Guanine
5. Cytosine 5. Cytosine
6. Uracyl 6. Thymine

The main difference between the two types of nucleic acid can thus be seen. In addition to the difference
in their sugar component, they are distinguisehed by the pyrimidine bases which they contain. While both
contain cytosine, the uracyl of RNA is replaced by thymine in DNA.

The two types are both found in plants and animals. RNA is largely found in the cytoplasm while DNA
occurs abundantly in the nucleus. Protein synthesis appears to be the special function of RNA molecules , while
DNA functions in the replication and transmission of specific genetic units, hence in heredity. It seems to be
definitely established now that no living cell can exist unless it possesses both types of nucleic acids. This
explains why viruses which are made up only of one type of nucleic acid (plant viruses have RNA and bacterial
or animal viruses have DNA) must associate with other living cells to provide for their reproduction, hence they
cause the infectious diseases in plants and animals.

RNA have been identified histologically by the orcinal color reactions for pentoses.
DNA gives the Feulgen’s reaction, which is generally believed to be specific for deoxyribose nucleic
acid.

Nomenclature of the nucleic acid units: the nucleotides are named after the purine or pyrimidine base
they contain thus:
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 N – base Nucleosides (with sugar) Nucleotides (nucleic acids)

Adenine Adenosine Adenylic acid

Guanine Guanosine Guanylic acid
Cytosine Cytidine Cytidylic acid
Thymine Thymidine Thymidylic acid
Uracyl Uridine Uridylic acid

The structural components of the nucleic acid maybe seen from below:

A. Sugar Components:

B. Nitrogenous Bases

1. Purine Bases

2. Pyrimidine Bases

Nucleotides of Biological Importance

Several nucleotides other than those serving as components of nucleic acids are found in tissues.
They perform important functions in the body. Among them are:

1. Adenine Derivatives : Three phosphate derivatives are well known : Adenosine

Monophosphate or Muscle Adenylic Acid (AMP), Adenosine Diphosphate (ADP) and

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Adenosine Triphosphate (ATP) are all involved in energy production in livng cells
by acting as carriers or transfer agents for the phosphate groups.

2. Uracyl Derivatives: Uridine Diphosphate Glucose (UDPG) and Uridine

Diphosphate Galactose (UDPGal) are involved in glycogen formation. Uridine
Triphosphate (UTP) is associated with energy transfer in the conversion of
glucose and galactose.

3. Cytosine Derivatives: Cytosine Triphosphate (CTP) has been found to be important

in the synthesis of phospholipid (especially lecithin and cephalin) in the body.

4. Coenzyme – Vitamin Substances : several vitamins of the B – complex group are

found in nucleotide linkages they function as co – enzymes in many biological
reactions of living cells.

a. Flavin Adenine Dinucleotide (FAD) and Flavin Mononucleotide (FMN).

Riboflavin or Vitamin B2 is the component vitamin.

b. Coenzyme 1 (Diphosphopyridine Nucleotide or DPN) and C[oenzyme II

(Triphosphopyridine Nucleotide or TPN). Niacin or nicotinic acid form an
integral part of the molecule.

c. Coenzyme A (Coacetylase) with pantothenic acid as the vitamin. It is

involved in the transacylation reactions in biological oxidation.

d. Vitamin B12 (-Cobamide) coenzyme; is found to be an important compound in

the synthesis of porphyrins (substances to which hemoglobin, cytochromes,
catalase and myoglobin belong).

e. Thiamine (as co – carboxylase and pyridoxine (co – transaminase) when

phosphorylated function as integral parts of co – enzyme involved also in
biological oxidation. Thiamine is 2,5 – dimethyl – 6 - amino

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pyrimidine. Pyridoxine is 2 – methyl – 3 – hydroxy – 4 – carboxy – 5 –
hydroxymethyl pyridi

Fig. 1 Structures of DNA and RNA

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