PROTEIN METABOLISM

MTY1109 Lecture

Department ofDepartment
Medical Technology,
of MedicalBiochemistry
Technology, Biochemistry
Cluster – 2021-2022
Cluster – 2021-2022
LESSON OUTLINE
1. Protein Digestion
2. Amino Acid Utilization
3. Transamination and oxidative deamination
4. The Urea Cycle
5. Glucogenic and Ketogenic Amino Acids

Department of Medical Technology, Biochemistry Cluster – 2021-2022

 OVERVIEW

Amino acids are not stored by the body

(unlike fats and carbohydrates)

Amino acids must be obtained from the diet, synthesized

de novo or produced from normal protein degradation

Ammonia excreted in the

1st phase of AA Removal of α- Formation of ammonia and
urine or used to synthesize
catabolism amino groups corresponding α-keto acid
urea

2nd phase of AA common intermediates of energy producing metabolic pathways

α-keto acids
catabolism (can be metabolized to CO2 and water, glucose, fatty acids, or ketone bodies)

Department of Medical Technology, Biochemistry Cluster – 2021-2022


LESSON OUTLINE
1. Protein Digestion
2. Amino Acid Utilization
3. Transamination and
oxidative deamination
4. The Urea Cycle
5. Glucogenic and
Ketogenic Amino Acids
Protein Digestion
• Protein digestion starts in the stomach
• Dietary protein present in the stomach
stimulates the release of gastrin
• Gastrin: promotes secretion of pepsinogen
and HCl (has 3 functions)
1. Antiseptic properties kill most bacteria
2. Denaturing action “unwinds” globular
proteins
3. Acidic property leads to activation of
pepsinogen
Department of Medical Technology, Biochemistry Cluster – 2021-2022
Protein Digestion
Pepsin affects the hydrolysis of 10% peptide
bonds
• Production of secretin is stimulated by the passage
of small amounts of acidic protein content into the
small intestine
• Secretin stimulates bicarbonate (HCO3-)
production, which in turn helps neutralize
acidified gastric content
• Promotes secretion of pancreatic digestive
enzymes trypsin, chymotrypsin, and
carboxypeptidase in their inactive forms

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Proteolytic Enzymes
(which also have their zymogen forms)

Enzymes Specific to
Pepsin (pH 1.3) N- and C-terminal cleavage of Phe and Leu
Trypsin C-terminal cleavage of Lys and Arg
Chymotrypsin C-terminal cleavage of aromatic amino acids

N terminal side - <amino acid residues> - C-terminal side

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Protein Digestion
• Liberated amino acids are transported into the bloodstream via active transport process
• The passage of polypeptides and small proteins across the intestinal wall is uncommon in adults
• In infants, the transport of polypeptides allows the passage of proteins such as antibodies in
colostrum milk from a mother to a nursing infant to build up immunologic protection in the
infant

Department of Medical Technology, Biochemistry Cluster – 2021-2022

 Amino Acid Utilization
• The Amino Acid Pool - total supply of free amino acids available for use in
LESSON OUTLINE the human body
1. Protein Digestion
2. Amino Acid
Utilization SOURCES ROUTES OF DEPLETION
3. Transamination and
oxidative deamination
4. The Urea Cycle 1. Synthesis of body protein
5. Glucogenic and
1. Amino acid degraded of body 2. AA consumed for the synthesis of
Ketogenic Amino Acids proteins essential nitrogen-containing
2. Amino acids derived from dietary molecules
protein 3. Conversion of AA to glucose,
3. Synthesis of non-essential AA from glycogen, fatty acids, ketone
intermediates of metabolism bodies, or H2O + CO2

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Protein Turnover
• The repetitive process in which proteins are degraded and
resynthesized.
• The rate of protein synthesis is just sufficient to replace the protein being
degraded
• Leads to hydrolysis and resynthesis of 300-400 g of body protein
each day

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Biosynthesis of non-essential amino acids
in the liver
• Nitrogen balance: the state that results when the amount of nitrogen taken into the
human body as protein equals the amount of nitrogen excreted from the body in waste
materials
• Negative nitrogen imbalance - Protein degradation exceeds protein synthesis
• Amount of nitrogen in urine exceeds consumed amount
• Results in tissue wasting
• Positive nitrogen imbalance - Rate of protein synthesis (anabolism) is more than
protein degradation (catabolism)
• Indicated by the synthesis of large amounts of tissue

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Uses for Amino Acids
Synthesis of non-protein
Synthesis of non-
Protein synthesis nitrogen-containing Production of energy
essential amino acids
compounds

• Essential amino acids

• Synthesis of purines
cannot be synthesized • Amino acids are not
• Uses approximately and pyrimidines
due to the lack of an stored in the body
75% of free amino acids • Synthesis of heme for
appropriate carbon (excess are degraded)*
hemoglobin
chain

* Each amino acid has a unique degradation pathway

* The amino nitrogen atom is removed and excreted from the body as urea
* The remaining carbon skeleton is converted to pyruvate, acetyl CoA, or a citric acid cycle intermediate

Department of Medical Technology, Biochemistry Cluster – 2021-2022


LESSON OUTLINE
1. Protein Digestion
2. Amino Acid Utilization
3. Transamination and
oxidative deamination
4. The Urea Cycle
5. Glucogenic and
Ketogenic Amino Acids
Transamination and Oxidative
Deamination
• Two stages of amino acid degradation:
• Removal of the α-amino group
• Degradation of the remaining carbon skeleton
• Glutamate Production via Transamination
• Glutamate: produced through transamination when α-ketoglutarate is
the amino group acceptor.
• Aminotransferase always utilized glutamine in its amino
transfers, as many amino acids have their respective keto-acid
equivalent which are derived from or are themselves Krebs cycle
intermediates
Department of Medical Technology, Biochemistry Cluster – 2021-2022
Transamination

Reactions catalyzed during

amino acid catabolism.
A. Alanine aminotransferase
(ALT) – transfers an amino
group from alanine
B. Aspartate aminotransferase
(AST) – transfers an amino
group from aspartate

Aminotransferase reaction using α-

ketoglutarate as the amino group acceptor.

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Department of Medical Technology, Biochemistry Cluster – 2021-2022
Oxidative Deamination
• Deamination - a biochemical reaction in which an α-
amino acid is converted to an α-keto acid with release
of an ammonium ion that occurs in the mitochondria
of the liver and kidney

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Oxidative Deamination

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Transport of Ammonia to the
Liver
1. Tissues use glutamine synthase to combine ammonia with
glutamate to form glutamine
2. Glutamine is then transported in the blood to the liver
3. Cleavage is then done by glutaminase to produce glutamate
and free ammonia

• IN THE MUSCLE
• Alanine is transported by the blood to the liver where it is
converted to pyruvate

Department of Medical Technology, Biochemistry Cluster – 2021-2022


LESSON OUTLINE
1. Protein Digestion
2. Amino Acid Utilization
3. Transamination and
oxidative deamination
4. The Urea Cycle
5. Glucogenic and
Ketogenic Amino Acids
The Urea Cycle
• Urea Cycle: the body’s only way to SAFELY remove ammonia
(NH3) from the body
• Ammonia build up can be toxic, hence the need for its
removal
• This is mainly described as a series of biochemical reactions
in which urea is produced from ammonium ions and
aspartate as nitrogen sources
• Urea: produced in the liver is transported via blood to
the kidneys and eliminated from the body in urine.
Department of Medical Technology, Biochemistry Cluster – 2021-2022
The Urea Cycle
1. CARBOMOYL GROUP TRANSFER
• Carbamoyl Phosphate: one of the sources
of fuel for the urea cycle
• Two ATP molecules are expended in
the formation of one carbamoyl
phosphate molecule. It contains a
high-energy phosphate bond and is
formed in the mitochondrial matrix.
• The carbamoyl group of carbamoyl
phosphate is transferred to ornithine to
form citrulline

Department of Medical Technology, Biochemistry Cluster – 2021-2022

The Urea Cycle
2. CITRULLINE-ASPARTATE CONDENSATION
• Citrulline is transported into the cytosol and reacts with aspartate to produce
argininosuccinate synthetase, utilizing ATP

Department of Medical Technology, Biochemistry Cluster – 2021-2022

The Urea Cycle
3. ARGININOSUCCINATE CLEAVAGE
• Argininosuccinate is cleaved to arginine and fumarate by the enzyme argininosuccinate
lyase

Department of Medical Technology, Biochemistry Cluster – 2021-2022

The Urea Cycle
4. UREA FROM ARGININE HYDROLYSIS
a) Hydrolysis of arginine produces urea
and regenerates ornithine under the
influence of arginase
b) The oxygen atom present in urea
comes from water
c) Ornithine is transported back to
mitochondria to be used in the urea
cycle

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Net Reaction
• Four (4) ATP molecules used in
the production of one urea molecule
• Two molecules of ATP are
consumed in the production of
carbamoyl phosphate
• The equivalent of two ATP
molecules is consumed in step
two of the urea cycle to give
AMP and the PPi

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Relationship of
the Urea Cycle
and Krebs
Cycle
• Fumarate produced is
ultimately converted to
aspartate
• Aspartate re-enters the
urea cycle at step two

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Relationship of
the Urea Cycle
and Krebs
Cycle
• Fumarate produced is
ultimately converted to
aspartate
• Aspartate re-enters the
urea cycle at step two

Department of Medical Technology, Biochemistry Cluster – 2021-2022

 Glucogenic and Ketogenic
LESSON OUTLINE
Amino Acids
1. Protein Digestion
2. Amino Acid Utilization
3. Transamination and • Transamination/oxidative deamination removes the amino group
oxidative deamination from an amino acid
4. The Urea Cycle
5. Glucogenic and • An α-keto acid that contains the skeleton of the amino acid is produced
Ketogenic Amino • Each of the 20 amino acids undergo a different degradation
Acids
process
• Products formed are among a group of seven intermediates
• Four products are intermediates in the citric acid cycle
• Three products are pyruvate, acetyl CoA, and acetoacetyl CoA

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Glucogenic and Ketogenic Amino Acids
• The amino acids converted to citric acid cycle intermediates can serve as glucose precursors
• The amino acids converted to acetyl CoA or acetoacetyl-CoA can contribute to the formation of
fatty acids

Glucogenic amino acids Ketogenic amino acids

An amino acid that has a carbon- An amino acid that has a carbon-
containing degradation product that can containing degradation product that can
be used to produce glucose via be used to produce ketone bodies (only
gluconeogenesis Leu and Lys are purely ketogenic)

Department of Medical Technology, Biochemistry Cluster – 2021-2022

 Gluco and
Glucogenic Ketogenic
ketogenic
Alanine Tyrosine
Arginine*
Asparagine
Aspartate

Glucogenic Cysteine

Nonessential
Glutamate

and Ketogenic
Glutamine
Glycine
Histidine*

Amino Acids Proline

Serine

Methionine Isoleucine
Leucine

Essential
Threonine Phenylalanine
Lysine
Valine Tryptophan

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Fates of carbon
skeletons of amino
acids
Glucogenic amino acids are
shaded blue, and ketogenic amino acids
are shaded green

*participates in more than one degradation

pathway

Department of Medical Technology, Biochemistry Cluster – 2021-2022

Inter-
relationships
between
pathways

Department of Medical Technology, Biochemistry Cluster – 2021-2022

 Credits:
Biochemistry Cluster
Joanne Ancajas
Paolo Robert Bueno

Protein Metabolism Sharah Kristine Bonguit

Kathleen Mae Cruz
Calline Danica Gomez
Carlene Rome Ledesma
MTY1109 Lecture Marie Juneau Mallari
THE END Jessa Marie Natividad
Leslie Reynoso
Vanesa Segun
James Dominic Vergara
Erika Stephanie Viloria

Department ofDepartment
Medical Technology,
of MedicalBiochemistry
Technology, Biochemistry
Cluster – 2021-2022
Cluster – 2021-2022