Classification of Proteins

Proteins are important macromolecules of the cells, formed by the polymerization of amino acids
according to the sequence of genetic code in the mRNA. Proteins are the mode of expression of the
genetic information. They perform a variety of duties in the cells such as they act as the structural
components of cells, enzymes, hormones, pigments, storage proteins and some toxins in the cells. The
proteins are classified into many categories based on different criterions.

Criterion for the classification of proteins:

Ø Proteins are classified based on the following THREE criterions:

(I). Classification based on STRUCTURE of Protein

(II). Classification based on COMPOSITION of Protein

(III). Classification based on FUNCTIONS of Proteins

(I). Classification of Proteins based on the Structure of Proteins

Ø Based on the structure, proteins are classified into 3 groups.

(A). Fibrous Proteins

(B). Globular Proteins

(C). Intermediate Proteins

Ø They are linear (long fibrous) in shape.

Ø Secondary structure is the most important functional structure of fibrous proteins.

Ø Usually, these proteins do not have tertiary structures.

Ø Physically fibrous proteins are very tough and strong.

Ø They are insoluble in the water.

Ø Long parallel polypeptide chains cross linked at regular intervals.

Ø Fibrous proteins form long fibres or sheaths.

Ø Functions of fibrous proteins: perform the structural functions in the cells.

Ø Examples of fibrous proteins: Collagen, Myosin, Silk and Keratin.

(B). Globular Proteins

Ø Globular proteins are spherical or globular in shape.

Ø The polypeptide chain is tightly folded into spherical shapes.

Ø Physically they are soft than fibrous proteins.

Ø They are readily soluble in water.

Ø Most of the proteins in the cells belong to the category of globular proteins.

Ø Functions: Form enzymes, antibodies and some hormones.

Ø Example: Insulin, Haemoglobin, DNA Polymerase and RNA Polymerase

(C). Intermediate Proteins

Ø Their structure is intermediate to linear and globular structures.

Ø They are short and more or less linear shaped proteins

Ø Unlike fibrous proteins, they are soluble in water.

Ø Function: blood clotting proteins

Ø Example: Fibrinogen

Classification of Proteins based on Composition:

(A). Simple Proteins

(B). Conjugated Proteins

(A). Simple Proteins

Ø Simple proteins composed of ONLY amino acids.

Ø Proteins may be fibrous or globular.

Ø They possess relatively simple structural organization.

Ø Example: Collagen, Myosin, Insulin, Keratin

(B). Conjugated Proteins

Ø Conjugated proteins are complex proteins.

Ø They contain one or more non-amino acid components.

Ø Here the protein part is tightly or loosely bound to one or more non-protein part(s).
Ø The non-protein parts of these proteins are called prosthetic groups.

Ø The prosthetic group may be metal ions, carbohydrates, lipids, phosphoric acids, nucleic acids and FAD.

Ø The prosthetic group is essential for the biological functions of these proteins.

Ø Conjugated proteins are usually globular in shape and are soluble in water.

Ø Most of the enzymes are conjugated proteins.

Ø Based on the nature of prosthetic groups, the conjugated proteins are further classified as follows:

$ Phosphoprotein: Prosthetic group is phosphoric acid, Example- Casein of milk, Vitellin of egg yolk.

$ Glycoproteins: Prosthetic group is carbohydrates, Example – Most of the membrane proteins, Mucin
(component of saliva).

$ Nucleoprotein: Prosthetic group is nucleic acid, Example – proteins in chromosomes, structural

$ Chromoproteins: Prosthetic group is pigment or chrome, Example: Haemoglobin, Phytochrome and

$ Lipoproteins: Prosthetic group is Lipids, Example: Membrane proteins

$ Metalloproteins: Prosthetic group is Metal ions, Example: Nitrate Reductase.

(III). Classification of Protein based on Functions:

(A). Structural Proteins:

Ø Form the component of the connective tissue, bone, tendons, cartilage, skin, feathers, nail, hairs and
horn.

Ø Most of them are fibrous proteins and are insoluble in water.

Ø Example: Collagen, Keratin and Elastin.

(B). Enzymes:

Ø They are the biological catalysts.

Ø Enzymes reduce the activation energy of reactants and speed-up the metabolic reactions in the cells.

Ø Most of them are globular conjugated proteins

(c). Hormones:

Ø They include the proteinaceous hormones in the cells.

Ø Example: Insulin, Glucagon, ACH

(D). Respiratory Pigments

Ø They are coloured proteins

Ø All of them are conjugated proteins and they contain pigments (chrome) as their prosthetic group.

Ø Example: Haemoglobin, Myoglobin

(E). Transport Proteins

Ø They transport the materials in the cells

Ø They form channels in the plasma membrane

Ø They also form one of the components of blood and lymph in animals.

Ø Example: Serum albumin

Ø They are the force generators of muscles

Ø They can contract with the expense of energy from ATP molecules.

Ø Example: Actin, Myosin

(G). Storage Proteins

Ø They act as the store of metal ions and amino acids in the cells.

Ø Found in seeds, egg and milk

Ø Abundantly seen in pulses (legume seeds).

Ø Example: Ferritin which stores iron, Casein, Ovalbumin, Gluten of Wheat

(F). Toxins

Ø They are toxic proteins

Ø Example: Snake venom

Proteins may be defined as the high molecular weight mixed polymers of α-amino acids joined together
with peptide linkage (-CO-N H-). Proteins are the chief constituents of all living matter. They contain
carbon, hydrogen, nitrogen and sulphur and some contain phosphorus also.

2. Biological Importance of Proteins:

ADVERTISEMENTS:

i. Proteins are the essence of life processes.

ii. They are the fundamental constituents of all protoplasm and are involved in the structure of the living
cell and in its function.

iii. Enzymes are made up of proteins.

iv. Many of the hormones are proteins.

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v. The cement substances and the reticulum which bind or hold the cells as tissues or organs are made
up partly of proteins.
vi. They execute their activities in the transport of oxygen and carbon dioxide by hemoglobin and special
enzymes in the red cells.

vii. They function in the homostatic control of the volume of the circulating blood and that of the
interstitial fluids through the plasma proteins.

viii. They are involved in blood clotting through thrombin, fibrinogen and other protein factors.

ix. They act as the defence against infections by means of protein antibodies.

x. They perform hereditary transmission by nucleoproteins of the cell nucleus.

3. Classification of Proteins:

I. Simple proteins

(i) Albumins:

Soluble in water, coagulable by heat and 1 precipitated at high salt concentrations.

ADVERTISEMENTS:

Examples – Serum albumin, egg albumin, lactalbumin (Milk), leucosin (wheat), legumelin (soyabeans).
(ii) Globulins:

Insoluble in water, soluble in dilute salt 1 solutions and precipitated by half 1 saturated salt solutions.

Examples – Serum globulin, vitellin (egg yolk), tuberin (potato), myosinogen (muscle), legumin (peas).

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(iii) Glutelins:

Insoluble in water but soluble in dilute 1 acids and alkalis. Mostly found in plants.

Examples – Glutenin (wheat), oryzenin (rice).

(iv) Prolamines: Insoluble in water and absolute alcohol 1 but soluble in 70 to 80 per cent alcohol.

ADVERTISEMENTS:

Examples – Gliadin (wheat), zein (maize).

Basic proteins of low molecular weight. 1 Soluble in water, dilute acids and alkalis, j Not coagulable by
heat.

Examples – Salmine (salmon sperm).

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(vi) Histones:

Soluble in water and insoluble in very I dilute ammonium hydroxide.

Examples – Globin of hemoglobin and thymus histones.

(vii) Scleroproteins:

Insoluble in water, dilute acids and alkalis.

Examples – Keratin (hair, horn, nail, hoof and feathers), collagen (bone, skin), elastin (ligament).

II. Conjugated Proteins

(i) Nucleoproteins:
Composed of simple basic proteins (protamines or histones) with nucleic acids, I found in nuclei. Soluble
in water.

Examples – Nucleoprotamines and nucleohistones.

(ii) Lipoproteins:

Combination of proteins with lipids, such ‘ as fatty acids, cholesterol and 1 phospholipids etc.

Examples – Lipoproteins of egg-yolk, milk and cell membranes, lipoproteins of blood.

(iii) Glycoproteins:

Combination of proteins with carbohydrate (mucopolysaccharides).

Examples – Mucin (saliva), ovomucoid (egg white), osseomucoid (bone), tendomucoid (tendon).

(iv) Phosphoproteins:

Contain phosphorus radical as a | prosthetic group.

Examples – Caseinogen (milk), ovovitellin (egg yolk).

(v) Metalloproteins:
Contain metal ions as their prosthetic | groups. The metal ions generally are Fe, I Co. Mg, Mn, Zn, Cu etc.

Examples – Siderophilin (Fe), ceruloplasmin (Cu).

(vi) Chromoproteins:

Contain porphyrin (with a metal ion) as | their prosthetic groups.

Examples – Haemoglobin , myoglobin, catalase, peroxidase, cytochromes.

(vii) Flavoproteins:

Contain riboflavin as their prosthetic 1 groups.

Examples – Flavoproteins of liver and kidney.

III. Derived Protein

A. Primary derivatives

(i) Proteans:

Derived in the early stage of protein hydrolysis by dilute acids, enzymes or alkalis.

Examp
Derived Protein

A. Primary derivatives

(i) Proteans:

Derived in the early stage of protein hydrolysis by dilute acids, enzymes or alkalis.

Examples – Fibrin from fibrinogen.

(ii) Metaproteins:

Derived in the later stage of protein hydrolysis by slightly stronger acids and alkalis.

Examples – Acid and alkali metaproteins.

(iii) Coagulated:

They are denatured proteins formed by the action of heat. X-rays, ultraviolet rays etc.

Cooked proteins, coagulated albumins.

B. Secondary derivatives

(i) Proteoses:
Formed by the action of pepsin or trypsin. Precipitated by saturated solution of ammonium sulphate,
incoagulable by heat.

Examples – Albumose from albumin, globulose from globulin.

(ii) Peptones: .

Further stage of cleavage than the proteoses. Soluble in water, incoagulable by heat and not precipitated
by saturated ammonium sulphate solutions.

(iii) Peptides:

Compounds containing two or more amino acids. They may be di-, tri-, and porypeptides.

Examples – Glycyl-alanine, leucyl-glutamic acid.

Physical Properties of Proteins

Colour and Taste

Proteins are colourless and usually tasteless. These are homogeneous and crystalline.

Shape and Size

The proteins range in shape from simple crystalloid spherical structures to long fibrillar structures. Two
distinct patterns of shape

have been recognized :

A. Globular proteins- These are spherical in shape and occur mainly in plants, esp., in seeds and in leaf
cells. These are bundles formed by folding and crumpling of protein chains. e.g., pepsin, edestin, insulin,
ribonuclease etc.
B. Fibrillar proteins- These are thread-like or ellipsoidal in shape and occur generally in animal muscles.
Most of the studies regarding protein structure have been conducted using these proteins. e.g.,
fibrinogen, myosin etc.

Molecular Weight

The proteins generally have large molecular weights ranging between 5 × 103 and 1 × 106. It might be
noted that the values of molecular weights of many proteins lie close to or multiples of 35,000 and
70,000.

Colloidal Nature

Because of their giant size, the proteins exhibit many colloidal properties, such as; Their diffusion rates
are extremely slow and they may produce considerable light-scattering in solution, thus resulting in
visible turbidity (Tyndall effect).

Denaturation

Denaturation refers to the changes in the properties of a protein. In other words, it is the loss of biologic
activity. In many instances the process of denaturation is followed by coagulation— a process where
denatured protein molecules tend to form large aggregates and to precipitate from solution.

Amphoteric Nature

Like amino acids, the proteins are amphoteric, i.e., they act as acids and alkalies both. These migrate in
an electric field and the direction of migration depends upon the net charge possessed by the molecule.
The net charge is influenced by the pH value. Each protein has a fixed value of isoelectric point (pl) at
which it will move in an electric field.

Ion Binding Capacity

The proteins can form salts with both cations and anions based on their net charge.

Solubility

The solubility of proteins is influenced by pH. Solubility is lowest at isoelectric point and increases with
increasing acidity or alkalinity. This is because when the protein molecules exist as either cations or
anions, repulsive forces between ions are high, since all the molecules possess excess charges of the
same sign. Thus, they will be more soluble than in the isoelectric state.

Optical Activity

All protein solutions rotate the plane of polarized light to the left, i.e., these are levoratotory.

Chemical Properties of Proteins

Proteins are hydrolyzed by a variety of hydrolytic agents.

A. By acidic agents: Proteins, upon hydrolysis with conc. HCl (6–12N) at 100–110°C for 6 to 20 hrs, yield
amino acids in the form of their hydrochlorides.

B. By alkaline agents: Proteins may also be hydrolyzed with 2N NaOH.

Reactions involving COOH Group

A. Reaction with alkalies (Salt formation)

B. Reaction with alcohols (Esterification)

C. Reaction with amines

Reactions involving NH2 Group

A. Reaction with mineral acids (Salt formation): When either free amino acids or proteins are treated
with mineral acids like HCl, the acid salts are formed.

B. Reaction with formaldehyde: With formaldehyde, the hydroxy-methyl derivatives are formed.

C. Reaction with benzaldehyde: Schiff ‘s bases are formed

D. Reaction with nitrous acid (Van Slyke reaction): The amino acids react with HNO2 to liberate N2 gas
and to produce the corresponding α-hydroxy acids.

E. Reaction with acylating agents (Acylation)

F. Reaction with FDNB or Sanger’s reagent

G. Reaction with dansyl chloride

Reactions involving both COOH AND NH2 Group

A. Reaction with triketohydrindene hydrate (Ninhydrin reaction)

B. Reaction with phenyl isocyanate: With phenyl isocyanate, hydantoic acid is formed which in turn can
be converted to hydantoin.

C. Reaction with phenyl isothiocyanate or Edman reagent

D. Reaction with phosgene: With phosgene, N-carboxyanhydride is formed

E. Reaction with carbon disulfide: With carbon disulfide, 2-thio-5-thiozolidone is produced

Reactions involving R Group or Side Chain

B. Xanthoproteic test

C. Millon’s test

D. Folin’s test

E. Sakaguchi test

F. Pauly test

G. Ehrlich test

Reactions involving SH Group

A. Nitroprusside test: Red colour develops with sodium nitroprusside in dilute [Link]. The test is
specific for cysteine.

B. Sullivan test: Cysteine develops red colour in the presence of sodium 1, 2-naphthoquinone- 4-
sulfonate and sodium hydrosulfite.

Properties of Proteins:

1. Denaturation:

Partial or complete unfolding of the native (natural) conformation of the polypeptide chain is known as
denaturation. This is caused by heat, acids, alkalies, alcohol, acetone, urea, beta- mercaptoethanol.

2. Coagulation:

When proteins are denatured by heat, they form insoluble aggregates known as coagulum. All the
proteins are not heat coagulable, only a few like the albumins, globulins are heat coagulable.

3. Isoelectric pH (pH1):
The pH at which a protein has equal number of positive and negative charges is known as isoelectric pH.
When subjected to an electric field the proteins do not move either towards anode or cathode, hence
this property is used to isolate proteins. The proteins become least soluble at pHI and get precipitated.
The pHI of casein is 4.5 and at this pH the casein in milk curdles producing the curd.

4. Molecular Weights of Proteins:

The average molecular weight of an amino acid is taken to be 110. The total number of amino acids in a
protein multiplied by 110 gives the approximate molecular weight of that protein. Different proteins
have different amino acid composition and hence their molecular weights differ. The molecular weights
of proteins range from 5000 to 109 Daltons. Experimentally the molecular weight can be determined by
methods like gel filtration, PAGE, ultra centrifugation or viscosity measurements