Tertiary Structure of

Proteins - Protein
Denaturation
GROUP 2
 ANALOGY:
Relationships among the primary,
Tertiary secondary, and tertiary structures
of a protein
Structure of • The primary structure is
Proteins
• is the overall three-
the long, straight cord
dimensional shape of a
protein that results from the • The secondary structure
interactions between amino is the coiling of the cord
acid side chains (R groups) into a helical arrangement
that are widely separated • The tertiary structure is the
from each other within a supercoiling arrangement the cord
peptide chain adopts after the receiver has been
hung up a number of times
 Interactions Responsible for Tertiary
Structure
Four types of stabilizing interactions contribute to the
tertiary structure of a protein:
1. Covalent disulfide bonds,
2. Electrostatic attractions (salt bridges)
3. Hydrogen bonds
4. Hydrophobic attractions.
All four of these interactions are interactions between amino acid R groups.
This is a major distinction between tertiary-structure interactions and secondary-structure
interactions:
• Tertiary-structure interactions involve the R groups of amino acids;
• Secondary-structure interactions involve the peptide linkages between amino acid
residues.
Interactions Responsible for Tertiary
Structure
Covalent Disulfide bond
Disulfide bonds
• the strongest of the tertiary-structure
interactions,
• Covalent interactions formed between
the sulfur atoms of two cysteine
residues.
• stabilize monomeric and multisubunit
proteins
Cysteine is the only a-amino acid that contains a sulfhydryl
Interactions Responsible for Tertiary
Structure
Electrostatic Interactions (salt bridges)
• always involve the interaction between an
acidic side chain (R group) and a basic side
chain (R group).
• When negatively charged atom is attracted
towards positively charged atom and vice-
versa
• The interaction that occurs between the two
types of side chains is a positive-negative ion-
ion attraction.
Interactions Responsible for Tertiary
Structure
Hydrogen bonds
• can occur between amino acids with polar R groups. A
variety of polar side chains can be involved, especially
those thal possess the following functional groups:

• relatively weak and are easily disrupted by changes in pH

and temperature.
 Interactions Responsible for Tertiary
Structure
Hydrophobic interactions
• result when two nonpolar side chains are close to each other.
• a kind of property of nonpolar molecules (or hydrophobic moieties of
amphiphiles), which can drive these molecules to assemble to form anhydrous
domains in aqueous solution
• are common between phenyl rings and alkyl side chains.
• Although hydrophobic interactions are weaker than hydrogen bonds or
electrostatic interactions, they are a significant force in some proteins because
there are so many of them: their cumulative effect can be greater in magnitude
than the effects of hydrogen bonding.
Disulfide bonds, electrost atic interactions, hydrogen bonds, and
hydrophobic interactions are all stabilizing influences that
contribute to the tertiary structure of a protein
Myoglobin Tertiary Structure
• In 1959, a protein tertiary structure was
determined for the first time.
• The determination involved myoglobin, a
conjugated protein whose function is oxygen
storage in muscle tissue.
• Structure:
• It involves a single peptide chain of 153
amino acids with numerous a helix segments
within the chain.
• a prosthetic heme group
• an iron-containing group with the ability to
bind molecular oxygen.
Quaternary Structure of Proteins
• Quaternary structure is the highest level of protein
organization.
• It is found only in multimeric proteins
• Such proteins have structures involving two or more
peptide subunits that are independent of each other—
that is, are not covalently binded to each other.
• Quaternary protein structure is the organization among the
various peptide subunits in a multimeric protein.
• Most multimeric proteins contain an even number of subunits
(two subunits = a dimer, four subunits = a tetramer, and so on)
• The subunits are held together by the same types of noncovalent
interactions that contribute to tertiary structure
(electro-static interactions, hydrogen bonds, and hydrophobic
tertiary and
interactions), quaternary
• Noncovalent interactions that contribute to quaternary structure, structure of
the oxygen-
are, however, weaker and thus more easily disrupted carrying
• It is significant that the different parts of a multimeric protein are protein
hemoglobin.
called protein subunits rather than protein chains. Covalent
bonding does not occur between protein subunits but can occur
between protein chains. (disulfide bonds).
Protein Structure
Protein Hydrolysis
• When a protein or smaller peptide in a solution of strong acid or strong base
is heated, the peptide bonds of the amino acid chain are hydrolyzed and free amino
acids are produced.
• The hydrolysis reaction is the reverse of the formation reaction for a peptide bond.
Amine and carboxylic acid functional groups are regenerated.
In COMPLETE protein hydrolysis:
• all peptide bonds are broken freeing up all of the constituent amino acids;
• amino acids are the only products.
In PARTIAL protein hydrolysis
• some, but not all, of the peptide bonds are broken producing a product mixture that
contains both free amino acids and small peptides.
 The complete hydrolysis of the tripeptide Ala-Gly-Cys under acidic
conditions produces one unit each of the amino acids alanine, glycine,
and cysteine.
• The equation for the hydrolysis is:

Protein digestion
- is simply enzyme-catalyzed hydrolysis of ingested protein.
- the hydrolysis of cellular proteins to amino acids is an ongoing process, as the
body resynthesizes needed molecules and tissue.
Protein denaturation
• is the partial or complete disorganization of a protein’s characteristic
three-dimensional shape as a result of disruption of its secondary,
tertiary, and quaternary structural interactions.
• Because the biochemical function of a protein depends on its three-
dimensional shape, the result of denaturation is loss of biochemical
activity.
• does not affect the primary structure of a protein.

Loss of water solubility

is a frequent physical consequence of protein denaturation
Protein Denaturation
Renaturation
• “Refolded” restoration process
• the reconstruction of a protein or nucleic acid (such as
DNA) to their original form especially after
denaturation.
• However, for extensive denaturation changes, the
process is usually irreversible.
Protein Denaturation
Coagulation
• precipitation out of biochemical solution of
denatured protein.
• example of protein denaturation occurs when egg
white (a concentrated solution of the protein
albumin) is poured onto a hot surface.
• The clear albumin solution immediately changes
into a white solid with a jellylike consistency
Protein Denaturation
Cauterization
• The process in which involves denaturation, where heat is often
used to seal small blood vessels in surgeries

Heat-induced Denaturation
• also used in sterilizing surgical instruments and in canning foods;
bacteria are destroyed when the heat denatures their protein.
Protein Denaturation
Alcohols
• are an important type of denaturing agent.
• Denaturation of bacterial protein takes place when isopropyl or ethyl alcohol is
used as a disinfectant-hence the common practice of swabbing the skin with
alcohol before giving an injection.
Pure Isopropyl or Ethyl Alcohol is less effective than the commonly used 70%
alcohol solution.
• Pure alcohol quickly denatures and coagulates the bacterial surface, thereby
forming an effective barrier to further penetration by the alcohol.
• The 70% solution denatures more slowly and allows complete penetration to be
achieved before congulation of the surface proteins takes place.