Biochemistry LEC

Protein Hydrolysis and Denaturation

OUTLINE

I. Protein Hydrolysis
II. Protein Denaturation
III. Processes in Amino Acid Degradation
IV. Disorders related to deficiencies with Protein

Hydrophilic and Hydrophobic Amino Acids

- Hydrophilic – amino acids that love water. Water loving amino acids.
- Hydrophobic – amino acids that fear water. Water fearing amino acids. They detest to water. If they are
subjected to water, they are most likely not to be hydrolyzed (meaning they stay as is, for they fear water).

Hydrophilic Hydrophobic
Arginine Alanine
Asparagine Isoleucine
Aspartic Acid Leucine
Cysteine Methionine
Glutamic Acid Phenylalanine
Glutamine Proline
Glycine Tryptophan
Histidine Tyrosine
Lysine Valine
Serine
Threonine

I. Protein Hydrolysis

- Peptide bonds broken through hydrolysis

- Hydrolysis
o Occurs in the stomach when enzymes catalyze the hydrolysis of proteins to give amino acids. Needed for
the metabolic processes in our body.
o Breaks up the primary structure by breaking covalent peptide bonds that link amino acids

Here are sequence of amino acids:

II. Protein Denaturation

- Occurs when the interactions of residues that stabilize the tertiary and quaternary structures are destructed
- Destroys the shape and renders the protein biologically inactive
- Does not necessarily mean with complete protein unfolding
- Occurs when a change disrupts the interactions between residues that stabilize the secondary, tertiary, or
quaternary structure
- Loss of secondary and tertiary in a protein occurs when conditions change such as:
o Increase in temperature
o Changing in ph
o Adding certain organic compounds or heavy metal ions
o Adding mechanical agitation
- When interactions between the residues are disrupted:
o A globular protein unfolds
o Tertiary structure is disrupted and the proteins is rendered biologically inactive

pH

- Changing the ph changes the charges on the side chains that disrupts electrostatic attractions and hydrogen
bonds
- Gastric acid in the stomach
 o If we intake proteins (Long chain of polypeptide), the enzymes in our stomach are responsible for
breaking down those proteins through their high acidic levels. Since ph is low, polar and nonpolar
structures of amino acids will be affected, so they will be breaking down making them inactive. Amino
acids will stay, and will be used in other processes of our body.

Organic Compounds

- Organic compounds such as ethanol and isopropyl alcohol acts as disinfectants by:
o Exchanging the bacterial protein’s hydrogen bonds to water
o Disrupting the side chain intramolecular hydrogen bonding
- Alcohol swab – used to clean wounds to prepare for skin puncture
o Best to use 70% concentration of alcohol because it enables to penetrate/denature the lipid bilayer of
bacterial cell wall. The bacterial cell wall is denatured. The remaining 30% (which is water) ay
magpuputok dun sa mismong bacteria to destroy it.
o If 100% mag dry lang si bacteria, tas pag pumasok yung tubig mabubuhay lang din yung bacteria

Heavy Metal Ions

- Heavy metals such as Ag+, Pb2+, and Hg2+

o Denature proteins by forming bonds with ionic residues or reacting with disulfide bonds
- Dilute (1%) AgNO3 “Silver Nitrate” – destroys gonorrhea infections in the eyes of the babies

Agitation

- Example: Beating egg whites

o When you beat eggs, they do not come back to their original form, meaning the whipping action
stretches the polypeptide chains. And when it is stretched, it will destabilize interactions with amino
acids.
- Whipping action stretched the polypeptide chains until the stabilizing interactions are disrupted.
Summary of Denaturation of Proteins

Denaturing Agent Bands disrupted Examples

Heat above 50C Hydrogen bonds, hydrophobic
interactions between nonpolar
residues
Changing in the pH Hydrogen bonds between poplar
residues, salt bridges
Organic Compounds Hydrophobic interaction
Heavy metal ions Disulfide bonds in proteins by
forming ionic bonds
Agitation Hydrogen and hydrophobic
interactions by stretching
polypeptide chains
Cooking food
Autoclaving medical instruments
Denaturation of milk protein from
bacteria (preparation of yogurt)
Ethanol and isopropyl alcohol for
disinfection
Silver Nitrate AgNO3
Beating of eggs

**When people are infected with prions, they most likely will develop mad cow disease. This disease is fatal and there is
not cure for it so far. Prions have a sequence that is complicated, which is why it is not reversible and it is very fatal.

III. Processes in Amino Acid Degradation

- 2 major enzyme systems: ATP-dependent Ubiquitin Proteasome (cytosol) and the ATP-independent
Degradative Enzyme System (lysosome)
- ATP-dependent Ubiquitin Proteasome System – modified by the covalent attachment of ubiquitin in eukaryotic
species
- ATP-independent Degradative Enzyme System – proteasomes selectively degrade damaged or short-lived
proteins, lysosomes uses hydrolases to selectively degrade intracellular and extracellular proteins
 - Ubiquitin (tigahatid sa protein substrate sa proteasome)
- Proteasome responsible to degradating protein substrate into different amino acids (or antigen presentation for
immunology)

Transamination difference to Protein Degradation

- Transamination is an exchange of a functional group between a keto acid. A process to convert your essential
amino acids to non-essential amino acids
- Oxidative deamination is the reverse of transamination

Amino acids go to degradation because those products will be involved in the metabolic processes (e.g. urea cycle) in
the body. (Carboxyl group, amine group, functional group, and the alpha carbons) Amine group involved in urea cycle
used in other metabolic processes in our body (E.g. Krebs cycle)

IV. Protein Deficiencies Disorders

 Marasmus
o Energy and protein deficiency and chronic energy deficit
o Characterized by edema, wasting, stunting, and mild hepatomegaly (bulging of stomach due to
enlargement of liver)
o Diseases associated with poverty
 Kwashiorkor
o Severe protein malnutrition and bilateral extremity swelling
o Relative protein deficiency but adequate energy intake
o Hypoproteinemia, pitting edema, dermatosis, and fatty infiltration of the liver
o Diseases associated with poverty
 Deficiencies of Protein C and Protein S
o Protein C and Protein S involved in blood coagulation (hematological disease)
 o Acts as natural anticoagulants or blood thinners
o High risk for pregnant women
 Cachexia
o Loss of weight, muscle atrophy, fatigue, weakness, and significant loss of appetite in someone who is not
actively trying to lose weight
o Cancer, AIDS, coeliac disease, COPD, multiple sclerosis, RA, CHF, tuberculosis, mercury poisoning, etc.
o More of a symptom rather than a disease itself.