Biochemistry for Medical Laboratory
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-
Protein Metabolism
TOPIC OUTLINE
1 Protein Digestion and Absorption
2 Amino Acid Utilization
3 Transamination and Oxidative Deamination
4 The Urea Cycle
5 Amino Acid Carbon Skeletons
6 Amino Acid Biosynthesis
7 Hemoglobin Catabolism
8 Interrelationship among Metabolic Pathways
Protein Digestion and Absorption
Protein digestion (denaturation and hydrolysis)
starts in the stomach rather than in the mouth
Reason: Saliva does don’t contain any enzyme that
affects the protein
 Denatured in the stomach by HCl
(hydrochloric acid-acidic pH)) in gastric juice
(pH of 1.5-2.0)
 Enzyme pepsin hydrolyzes about 10%
peptide bonds producing a variety of
polypeptide
Large polypeptide (partially digested proteins)
chains pass from stomach into small intestine
(where the digestion is completed):
 pH in small intestine is 7.0 -8.0 and helps
neutralize the acidified gastric content
 Trypsin, chymotrypsin and
carboxypeptidase (attack the peptide
bonds) in pancreatic juice released into the
small intestine help hydrolyze proteins to
smaller peptides
 Aminopeptidase secreted by intestinal
mucosal membrane further hydrolyze the
small peptides to amino acids
 Amino acids (aa) liberated are transported
into blood stream via active transport
process (requires ATP or energy in the
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
(made up of protein-large proteins)) in
colostral milk from a mother to a nursing infant to
build up immunologic protection in the infant.
 Proteolytic Enzymes (Trypsin, chymotrypsin
carboxypeptidase, and aminopeptidase) are
produced in inactive forms called zymogens
that are activated at their site of action.
 From an energy production standpoint, protein
supplies only a small portion of the body’s need.
 With the normal diet, carbohydrates and fats
supply 90% of the body’s energy and only 10%
are coming from the proteins.
 Protein metabolism plays an important role in
maintaining good health
 The amino acids obtained from protein synthesis
are needed for protein synthesis and synthesis
of other nitrogen containing compounds in
the cell.
Amino acids formed through digestion process
enters the amino acid pool in the body (counterpart
of the glycogen):
 Amino acid pool: the total supply of free amino
acids available for use in the human body
 Once a protein reaches your stomach,
hydrochloric acid enzymes breaks it down into
smaller chains of amino acids.
The amino acid pool is derived from 3 sources:
1. Dietary protein- proteins that are found in
our food sources; in the food that we ingest
2. Protein turnover: A repetitive process in
which the body proteins are degraded and
resynthesized (recycle the protein)
3. Biosynthesis of amino acids in the liver
o only non-essential amino acids are
synthesized
Dietary Protein
1. Mouth
o Saliva- no effect on digestion
2. Stomach
o HCl- denatures protein
o Pepsin- hydrolyzes peptide bonds
o Large Polypeptides
3. Small Intestine
o Hydrolyze peptide bonds
 Trypsin
 Chymotrypsin
 Carboxypeptidase
 Aminopeptidase
o Amino Acids
4. Intestinal Linings
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o Active transport  Synthesis of heme for hemoglobin,

o Amino acids in bloodstream neutrotransmitters and hormones

3. Synthesis of nonessential amino acids

Amino Acid Utilization
 Essential amino acids can’t be
Nitrogen Balance synthesized because of the lack of
appropriate carbon chain
 The state that results when the amount of
 Root black preventing the synthesis
nitrogen taken into the human body as protein
equals the amount of nitrogen excreted from the of the essential amino acid is not
body in waste materials. (adult) the lack of nitrogen but the lack of
correct carbon skeleton upon which
enzymes can work
Two types of nitrogen imbalance can occur in
 The essential amino acids
human body:
contain carbon chains or
Negative Nitrogen Positive Nitrogen
aromatic links not present in either
Imbalance Balance
amino acids or the intermediates of
 Protein  Rate of protein
carbohydrates or lipid metabolism
degradation synthesis
exceeds protein (anabolism) is
4. Production of energy
synthesis more than protein
 Amount of N in degradation  Amino acids are not stored in the
urine exceeds (catabolism) body, so the excess is degraded
nitrogen  Results in large  Each amino acid has a different
consumed amounts of tissue mechanism of degradation
 Results in tissue synthesis
wasting (because  During growth, Amino Acid Utilization
more tissue pregnancy and
proteins are being convalescence Degradation Pathways
catabolized that from emaciating  The amino nitrogen atom is removed and
are being replaced illness converted to ammonium ion, which ultimately is
by protein  The nitrogen intake excreted from the body as urea.
synthesis) exits the nitrogen
 Protein poor diet, output  The remaining carbon skeleton is then
starvation and converted to pyruvate, acetyl CoA, or a citric
wasting illnesses acid cycle intermediate, depending on its
produce a negative makeup, with the resulting energy production
balance or energy storage.

Amino Acid Utilization Transamination and Oxidative Deamination

Amino acids from the body's amino acid pool are used
in four different ways:
 Acetylcholine and Serotonin
1. Protein Synthesis
 About 75% of amino acids go into
synthesis of proteins that is needed
continuous replacement of old tissues
(protein turnover) and to build new
tissues (growth).
Degradation of an amino acid takes place in two
stages:
1. The removal of the -amino group and
2. The degradation of the remaining carbon
skeleton
Removal of amino group is a two-step process
1. Transamination- Biochemical process in
which the amino group of an alpha-amino
acid is transferred to an alpha-keto acid.

2. Synthesis of non-protein nitrogen- 2. Oxidative deamination- an amino acid is

containing compounds converted into the corresponding keto acid
 Synthesis of purines and pyrimidines by the removal of the amine functional group
for nucleic acid synthesis as ammonia and the ammonia eventually

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goes into the urea cycle.
Transamination: Involves transfer of the amino
group of an -amino acid to an alpha keto acid as
shown in the reaction below:
 Transamination is an enzyme catalyzed
reactions
 There are at least 50 transaminase
enzymes associated with transamination
reactions
 Most enzymes are specific for alpha-
ketoglutarate as the amino acid acceptor
but are less specific for the amino acid
 Glutamate is actually the amino acid
produced from the action of alpha-
ketoglutarate specific amino transferases
Initial effect of transamination: Collect the amino
groups from a variety of amino acids into just two
amino acids—glutamate (most cells) and alanine
(muscle cells)
 Transport to the liver, the alanine is then
converted to glutamate through
transamination reaction
 Glutamate then acts as the source of the
amino groups for continued nitrogen
metabolism or excretion or biosynthesis
 Aminotransferases are important to medical
technologist
 The concentration of amino transferases in
the blood is usually used to diagnose liver
and heart disorders
o Alanine amino transferase
o Aspartate amino transferase
 Net effect of transamination: Collection of
the amino groups from a variety of amino
acids into a single compound—the amino
acid glutamate
 To regenerate pyruvate and oxaloacetate for
use in further transamination reactions
 Ammonium ion (NH4+) group is liberated from
the glutamate amino acid formed from
transamination
 Oxidative deamination reaction is a
biochemical reaction catalyzed by glutamate
dehydrogenase in which glutamate is converted
into alpha-keto glutarate with the release of an
ammonium ion
 Occurs in liver and kidney
 The ammonium ion produced by oxidative
deamination is a toxic substance, so it is quickly
converted carbomoyl phosphate and then to
urea via the urea cycle/ornithine cycle in
mammals
 Two amino acids, serine and threonine,
undergo direct deamination by dehydration-
hydration process rather than oxidative
deamination
Urea Cycle
 The net effect of amino acid degradation is the
production of ammonium ion which is toxic and it
is converted to urea (relatively non-toxic
compound) in the liver via urea cycle.
 Urea cycle is a series of biochemical reactions in
which urea is produced from ammonium ions
and carbon dioxide.
 Urea is transported via the blood from liver to the
kidneys and eliminated from the body via urine.
 Ornithine Cycle- converts excess ammonia into
urea in the mitochondria of liver cells
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Urea:  The oxygen atom present in the urea

 white solid comes from water
 melting point 133 C
 very soluble in water Enzyme: Arginase
 odorless and colorless and has a salty taste
o does not contribute to the color or  Orthinine is transported back to
odor of urine mitochondria to be used in the urea
cycle
 Carbamoyl Phosphate is the fuel for the urea
cycle (formed from the oxidative deamination)
 Two ATP molecules are expended in the
formation of one carbamoyl phosphate molecule
 A high energy phosphate bond is present in
carbamoyl phosphate
 It takes place in mitochondrial matrix

Steps of the Urea Cycle

Stage 1: Carbomoyl group transfer

 The carbamoyl group of carbamoyl
phosphate is transferred to ornithine
to form citrulline

Enzyme: Ornithine
Transcarbamoylase

 Introduction of two nitrogen atoms and

carbon atom

Stage 2: Citrulline-aspartate condensation

 Citrulline is transported into the cytosol,
citrulline reacts with aspartate to
produce argininosuccinate utilizing Urea Cycle Net Reaction
ATP
 Total of four ATP molecules is expended in the
Enzyme: Argininosuccinate Synthase production of one urea cycle molecule

 In this reaction the second of two
nitrogen atoms of urea is introduced into
the cycle (One nitrogen comes from
carbamoyl phosphate and the other
from aspartate -- original source of both
is glutamate) collecting agent for amino
acid nitrogen
Stage 3: Argininosuccinate cleavage:
 Argininosuccinate is cleaved to
arginine and fumarate by the enzyme
argininosuccinate lyase
Stage 4: Hydrolysis of urea from arginine:
 Hydrolysis of arginine produces urea
and regenerates ornithine - one of the
cycle’s starting materials
o Two molecules are consumed in the
production of carbamoyl phosphate
and the equivalent of two ATP molecule
is consumed in step 2 of the urea cycle
to give AMP and two Pi
Linkage Between the Urea and Citric Acid
Cycles
 Fumarate produced is used in citric acid
cycle (net equation for urea formation)
o Entered citric acid cycle which is
converted to malate to oxaloacetate
to aspartate through transamination
 Aspartate produced through transamination
is used in the urea cycle at step 2

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Fates of Carbon Skeletons of Amino Acids

Function of oxaloacetate:

1. The oxaloacetate produced from fumarate of the

urea cycle can be converted to glucose via
gluconeogenesis
2. It can be condensed with acetyl coenzyme A to
form citrate
3. It can be converted to pyruvate
Amino Acid Carbon Skeletons
 Transamination and oxidative deamination
produces an alpha-keto acid that contains the
carbon skeleton from the amino acid
 Each of 20 amino acids carbon skeletons
undergo a different degradation process
 Degraded products are pyruvate, acetyl CoA
acetoacetyl CoA, alpha-ketoglutarate, succinyl
CoA, fumarate, and oxaloacetate
o Last four are intermediates in the
citric acid cycle
 Amino acid that can be degraded to pyruvate
can be either glucogenic or ketogenic
 Pyruvate can be metabolized to either
oxaloacetate or acetyl coenzyme A
 Metabolized the pyruvate as oxaloacetate
these are the glucogenic amino acid
 Pyruvate as coenzyme A these are the
ketogenic amino acid
Purely ketogenic amino acids:
o Leucine
o Lysine
Legend:
 Glucogenic is blue while ketogenic is green
 * have more than one degradation pathway and
present more than once in the diagram

The amino acids converted to citric acid cycle Amino Acid Biosynthesis
intermediates can serve as glucose precursors
(glucogenic amino acids).  Non-essential amino acids are synthesized in1-
3 steps
o Glucogenic amino acid: An amino acid  Essential amino acids are synthesized in 7-10
that has a carbon-containing steps
degradation product that can be used to  Excess amino acids are converted to fat and
produce glucose via stored
gluconeogenesis.
 Diet with lack of high-quality proteins results in
breakage of body proteins
The amino acids converted to acetyl CoA or
acetoacetyl CoA can serve as fatty acids and/or
ketone body precursors (ketogenic amino acids)

o Ketogenic amino acid: An amino acid

that has a carbon-containing
degradation product that can be used to
produce ketone bodies

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Summary of the Starting Materials for the
Biosynthesis of the 11 Nonessential Amino Acids
Starting Materials for the Biosynthesis of the 11
Nonessential Amino Acids
o 3-phosphoglycerase
o Pyruvate
o Oxaloacetate
o Alpha ketoglutarate
The non-essential amino acid are biosynthesized by
transamination of the appropriate alpha keto acid
material:
o Alanine
o Aspartase
o Glutamate
Tyrosine is obtained from the essential amino acid
phenyl alanine in a one-step oxidation
Hemoglobin Catabolism
 Red blood cells (RBCs) are highly specialized
cells whose primary function is to deliver
oxygen to cells and remove carbon dioxide
from body tissues
 Mature red blood cells have no nucleus or DNA
-- filled with red pigment hemoglobin
 Red blood cells are formed in the bone marrow
o 200 billion new red blood cells are
formed daily
 The life span of a red blood cell is about 4
months or 120 days
Hemoglobin is a conjugated protein with two
parts:
 Protein portion is globin
 Prosthetic group is heme
 Iron atom interacts with oxygen forming a
reversible complex (oxygen can come on and off)
with it
Old RBCs are broken down in the spleen (primary
site) and liver (secondary site):
Degradation of hemoglobin
 Globin protein part is converted to amino
acids and are put in amino acid pool
 Fe atom becomes part of ferritin -- an iron
storage protein -- saves the iron for use in
biosynthesis of new hemoglobin molecules
 The heme (tetrapyrrole) is degraded to bile
pigments and eliminated in feces or urine.
Bile Pigments
Bile pigments: The tetrapyrrole degradation products
secreted via the bile.
There are four bile pigments:
1. Biliverdin - green in color
2. Bilirubin - reddish orange in color.
3. Stercobilin –brownish in color (gives feces
their characteristic brown color).
4. Urobilin - yellow in color and present in
urine (gives characteristic yellow color to
urine).
 Daily normal excretion of bile pigments: 1–2
mg in urine and 250–350 mg in feces.
 Jaundice: Results from liver, spleen and
gallbladder malfunction.

o Results in higher than normal
bilirubin levels in the blood and gives
the skin and white of the eye yellow tint.
o Infectious hepatitis and cirrhosis
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Interrelationship among Metabolic Pathways

 The metabolic pathways of carbohydrates, lipids,

and proteins are integrally linked to one
another.
o A change in one pathway can affect
many other pathways.

Examples:
 Feasting (over-eating): Causes the body to
store a limited amount as glycogen and the
rest as fat.
 Fasting (no food ingestion): The body
uses its stored glycogen and fat for energy.
 Starvation (not eating for a prolonged
period):

o Glycogen stores are depleted,

o Body protein is broken down to amino
acids to synthesize glucose.
o Fats are converted to ketone bodies.

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