Wallaga University

Institute of Health Sciences

School of Medicine

Module name: Metabolic Homeostasis

Module code: MEHO2022

20-Feb-25 By Kejela B.(B. Pharm, MSc) 1

 Amino acid metabolism

By Kejela B. (B. Pharm, MSc.)

Lecture-7
20-Feb-25 By Kejela B.(B. Pharm, MSc) 2
 Amino acid pool
• The amount of free amino acids distributed throughout the
body is called amino acid pool
• Plasma level for most amino acids varies throughout the day
• It ranges between 4-8mg/dl.
• It tends to increase in the fed state and tends to decrease the
post absorptive state
• Sources of amino acid
– Dietary protein
– Breakdown of tissue proteins
– Biosynthesis of nonessential amino acids
20-Feb-25 By Kejela B.(B. Pharm, MSc) 3
 Protein turnover
• The amino acid pool within cells is generated from
– dietary amino acids
– degradation of proteins within the cell.
• Proteins are continuously synthesized and degraded in the
body.
• Examples of proteins that undergo extensive synthesis and
degradation are
– Hemoglobin
– muscle proteins
– digestive enzymes
– the proteins of cells sloughed off from GIT
20-Feb-25 By Kejela B.(B. Pharm, MSc) 4
 Protein turnover cont.
• Hemoglobin is produced in reticulocytes and reconverted to
amino acids by the phagocytic cells

• Muscle protein is degraded during periods of fasting and the

amino acids are used for gluconeogenesis

• After ingestion of protein muscle protein is resynthesized.

• A large amount of protein is recycled daily in the form of

digestive enzymes

• One-fourth of the cells lining the walls of the GIT are lost each
day and replaced by newly synthesized cells.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 5

 Protein turnover cont.
• As cells leave the GIT wall their proteins and other components
are digested by enzymes in the lumen of the gut

– Then the products are absorbed.

• Red blood cells have a life span of about 120 days.

• The hemoglobin in these cells is degraded to amino acids by

lysosomal proteases

– their amino acids are reused in the synthesis of new

proteins.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 6

 Protein turnover cont.
• 6% of the protein that enters the digestive tract is excreted in
the feces each day.

– dietary proteins, digestive enzymes and proteins in

sloughed-off cells

– The remainder is recycled.

• The differences in amino acid composition of body proteins

and recycling of amino acids are important factors

– to determine the requirements for specific amino acids

and total protein in the diet.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 7
 Protein turnover cont.
• The synthesis of many enzymes is induced in response to
physiological demand (fasting or feeding).

• These enzymes are continuously being degraded.

• Intracellular proteins are also damaged by oxidation and

other modifications.

• Mechanisms for intracellular degradation of unnecessary or

damaged proteins:

– involve lysosomes and the ubiquitin-proteasome

system.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 8
 Protein turnover cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 9

 Lysosomal protein turnover
• Lysosomes participate in the process of autophagy
– intracellular components are surrounded by membranes that
fuse with lysosomes and endocytosis
• Autophagy is complex regulated process in which cytoplasm is
sequestered into vesicles and delivered to the lysosomes.
• In lysosomes, the cathepsin family of proteases degrades the
ingested proteins to individual amino acids.
• The recycled amino acids can then leave the lysosome and
rejoin the intracellular amino acid pool.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 10

 Lysosomal protein turnover cont.

• The cathespins exhibit optimal activity within the acidic

environment of the lysosome.

• Starvation of a cell is a trigger to induce autophagy.

– allow old proteins to be recycled and the newly released

amino acids used for new protein synthesis

– to enable the cell to survive in starvation conditions.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 11

 The Ubiquitin-Proteasome Pathway

• Ubiquitin is small protein (76 amino acids) and highly

conserved.

• Ubiquitin targets intracellular proteins for degradation

– by covalently binding to the ε-amino group of lysine

residues.

– accomplished by three-enzyme system that adds ubiquitin

to proteins targeted for degradation.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 12

 The Ubiquitin-Proteasome Pathway cont.
• the target protein is polyubiquitinylated in which additional
ubiquitin molecules are added to previous ubiquitin molecules
– form a long ubiquitin tail on the target protein.
• After polyubiquitinylation is complete, the targeted protein is
released from the three-enzyme complex
– directed to the proteasome via a variety of mechanisms.
• A protease complex then degrades the targeted protein into
small peptides.
– release intact ubiquitin that can again mark other proteins
for degradation
20-Feb-25 By Kejela B.(B. Pharm, MSc) 13
 The Ubiquitin-Proteasome Pathway cont.
• Three enzymes involved:
1. E1, ubiqutiin activating protein
2. E2, Ubiquitin conjugating enzyme
3. E3, ubiquitin-protein ligase.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 14

 The Ubiquitin-Proteasome Pathway cont.
• The proteasome is cylindrical 20S protein complex with multiple
internal proteolytic sites.
• ATP hydrolysis is used both to unfold the tagged protein and
to push the protein into the core of cylinder.
• The complex is regulated by 19S regulatory particles which
bind the ubiquinylated protein and deliver them to the
complex.
• After the target protein is degraded, ubiquitin is released intact
and recycled.
– amino acids join the intracellular pool of free amino acids.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 15
 The Ubiquitin-Proteasome Pathway cont.

FIG. The proteasome and regulatory proteins

20-Feb-25 By Kejela B.(B. Pharm, MSc) 16

 Nitrogen balance
• the balance between protein synthesis and protein
degradation
• 100 grams of dietary protein contains approximately 16g of
nitrogen.
• Eighty-three percent of this ingested nitrogen eventually leaves
the body as urea
– 10% as ammonium ion
– 7% as organic waste products (uric acid and creatinine).
• 95% of the urea and a large majority of the other nitrogenous
wastes are excreted in the urine.
• The nitrogen balance is the difference between the nitrogen
entering and leaving the body. 17
 Nitrogen balance cont.
• A healthy adult with adequate protein intake should be in
nitrogen equilibrium
– amount of outgoing nitrogen matches the amount of
incoming nitrogen
– the amount of body protein remains constant.
 Positive nitrogen balance
• Observed when nitrogen intake exceeds nitrogen excretion
and increases amount of body protein
• Growing children, pregnant women, bodybuilders and
patients recovering from severe illnesses have a positive
nitrogen balance.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 18
 Nitrogen balance cont.

 Negative nitrogen balance

• Observed in dietary protein deficiency.

• Protein-starved body degrades 30-40g of amino acids

every day.

– This amount defines the dietary protein requirement.

• Essential amino acid deficiency has the same effect

– because protein synthesis is impaired

– even if only one of the essential amino acids is

missing.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 19
 Nitrogen balance cont.
• Patients with chronic infections, cancer or severe diseases
have a negative nitrogen balance

– because glucocorticoids and stress hormones favor

protein degradation

– thereby supplying amino acids for gluconeogenesis.

• Cytokines are biologically active proteins

– released by white blood cells in many diseases

– have catabolic effects on proteins like the stress

hormones.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 20
 Nitrogen balance cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 21

 Biosynthesis of the nonessential amino acids

20-Feb-25 By Kejela B.(B. Pharm, MSc) 22

 Biosynthesis of the nonessential amino acids
• Four amino acids are derived from intermediates of glycolysis:

– Serine

– Glycine

– cysteine

– alanine

• Six amino acids are derived from tricarboxylic acid (TCA) cycle
intermediates.

• Tyrosine is synthesized from essential amino acid phenylalanine

20-Feb-25 By Kejela B.(B. Pharm, MSc) 23

 Biosynthesis of the nonessential amino acids cont.

Fig: Overview of the synthesis of the nonessential amino acids

20-Feb-25 By Kejela B.(B. Pharm, MSc) 24

 Serine, glycine, cysteine and alanine

• Serine is synthesized from 3-phosphoglycerate

• Alanine is formed by transamination of pyruvate.

• Glycine is primarily produced from serine

• The major route of production of glycine from serine is by a

reversible reaction that involves FH4 and PLP

• The minor pathway for glycine formation involves threonine

degradation.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 25
 Serine, glycine, cysteine and alanine cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 26

 Serine, glycine, cysteine and alanine cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 27

 Serine, glycine, cysteine and alanine cont.
• The carbons and nitrogen for cysteine synthesis are provided
by serine
– sulfur is provided by methionine
• Serine reacts with homocysteine to form cystathionine.
– catalyzed by cystathionine β-synthase.
• Cleavage of cystathionine by cystathionase produces
cysteine and α-ketobutyrate
– forms succinyl CoA via propionyl CoA.
• Both cystathionine β-synthase (β-elimination) and
cystathionase (γ-elimination) require PLP.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 28
FIG. Synthesis and degradation of
cysteine

20-Feb-25 By Kejela B.(B. Pharm, MSc) 29

 Serine, glycine, cysteine and alanine cont.
• Cysteine inhibits cystathionine-β-synthase and regulates its
own production.

• Cysteine derives its sulfur from the essential amino acid

methionine

• Cysteine becomes essential if methionine is inadequate for

cysteine synthesis.

• adequate dietary source of cysteine spares methionine

– it decreases the endogenously degraded to produce

cysteine.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 30
 Glutamate, Glutamine, Proline, Arginine
Ornithine, Aspartate and Asparagine
• Two groups of amino acids are synthesized from TCA cycle
intermediates
– one group from α-ketoglutarate
– one from oxaloacetate
• Five carbons of glutamate are derived from α-ketoglutarate
– by transamination or glutamate dehydrogenase.
– B/c α-ketoglutarate can be synthesized from glucose
all of the carbons of glutamate can be obtained from
glucose
20-Feb-25 By Kejela B.(B. Pharm, MSc) 31
 Glutamate
• Glutamate is used for

– the synthesis of glutamine, proline, ornithine and arginine

– providing the glutamyl moiety of glutathione

γ-glutamyl-cysteinyl-glycine

20-Feb-25 By Kejela B.(B. Pharm, MSc) 32

 Glutamate cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 33

 Arginine
• Arginine is synthesized from glutamate via glutamate
semialdehyde to form ornithine
– The reactions of the urea cycle then produce arginine.

• However, arginine generated by urea cycle are adequate

only for the adult and are insufficient to support growth.
• Therefore, during periods of growth, arginine becomes an
essential amino acid.
• Arginine is cleaved by arginase to urea and ornithine.
• Excess ornithine is transaminated to glutamate semialdehyde
then oxidized to glutamate
20-Feb-25 By Kejela B.(B. Pharm, MSc) 34
 Arginine cont.

FIG. Synthesis and degradation of arginine

20-Feb-25 By Kejela B.(B. Pharm, MSc) 35
 Aspartate and Asparagine
• Aspartate is produced by transamination of oxaloacetate.
• This reaction is readily reversible
– so aspartate can be reconverted to oxaloacetate.
• Asparagine is formed from aspartate
– glutamine provides the nitrogen to form amide group.
• Asparaginase hydrolyzes asparagine to NH4+ and aspartate
• In the synthesis of glutamine from glutamate ammonium ion
(NH4+) provides the nitrogen.
• Glutaminase hydrolyzes glutamine to NH4+ and glutamate.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 36

 Tyrosine

• Phenylalanine is converted to tyrosine via hydroxylation

reaction

• The phenylalanine hydroxylase (PAH) requires molecular

oxygen and BH4.

• BH4 is synthesized in the body from guanosine triphosphate

(GTP).

• Dihydrobiopterin is converted to BH4 to produce tyrosine.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 37
 Tyrosine cont.

FIG. Hydroxylation of phenylalanine.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 38
 Amino acid degradation

20-Feb-25 By Kejela B.(B. Pharm, MSc) 39

 Amino acid degradation

FIG. Degradation of amino

acids.
A. Amino acids that produce
pyruvate or intermediates
of the TCA cycle.
B. Amino acids that produce
acetyl CoA or ketone
bodies.
20-Feb-25 40
 Glycine
• Glycine degradation can occur via two routes.

1. Conversion to CO2, NH3 and N5,N10-methylenetetrahydrofolate

– only pathway for glycine oxidation that can generate

energy

2. Conversion to glyoxylate by the enzyme D-amino acid oxidase

– oxidized to oxalate

– oxalate is sparingly soluble and precipitate in kidney tubules

 leading to kidney stone formation.

• About 40% of oxalate formed in the liver comes from glycine

metabolism.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 41
 Glycine cont.

FIG. The metabolism of glycine

20-Feb-25 By Kejela B.(B. Pharm, MSc) 42
 Glycine cont.
• The major reaction of glycine degradation in human tissues is
the glycine cleavage to CO2 and NH.
• A specialized metabolic product formed from glycine in
bacteria (but not humans) is trimethylamine

20-Feb-25 By Kejela B.(B. Pharm, MSc) 43

 Methionine
• Methionine is converted to S-adenosylmethionine (SAM)

– SAM donates its methyl group to other compounds to form

S-adenosylhomocysteine (SAH).

• SAH is then converted to homocysteine

• Methionine can be regenerated from homocysteine by a

reaction requiring both FH4 and vitamin B12

• Carbons of homocysteine are converted to α-ketobutyrate

– undergoes oxidative decarboxylation to propionyl CoA.

• The propionyl CoA is then converted to succinyl CoA

20-Feb-25 By Kejela B.(B. Pharm, MSc) 44
 Methionine cont.

FIG. Conversion of methionine and other amino acids to succinyl CoA.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 45
 Methionine cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 46

 Histidine
• Histidine is degraded to glutamate

20-Feb-25 By Kejela B.(B. Pharm, MSc) 47

 Proline, arginine and ornithine
• Proline, arginine and ornithine are degraded to glutamate

20-Feb-25 By Kejela B.(B. Pharm, MSc) 48

 Branched-chain amino acids
• Branched-chain amino acids are valine, isoleucine and leucine

• Branched-chain amino acids make up almost 25% of the

content of the average protein

• The degradative pathway for valine and isoleucine has two

major functions.

– for energy generation

– to provide precursor to replenish TCA cycle intermediates

• Both valine and isoleucine contain carbons that form succinyl

CoA.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 49
 Branched-chain amino acids cont.

• First, the branched-chain amino acids are transaminated.

• Second, α-keto analogues of these amino acids oxidatively

decarboxylated by the α-keto acid dehydrogenase complex

– Mechanism of reaction and cofactor requirements are

similar to

pyruvate dehydrogenase

α-ketoglutarate dehydrogenase.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 50

 Branched-chain amino acids cont.
• the highest level of activity for this dehydrogenase is found in

muscle tissue

• Like, β-oxidation of fatty acids NADH and FADH2 are

generated for energy production.

• Valine and isoleucine are converted to succinyl CoA

• Isoleucine also forms acetyl CoA.

• Leucine does not produce succinyl CoA.

• It forms acetoacetate and acetyl CoA and is ketogenic.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 51

 Branched-chain amino acids cont.

20-Feb-25 52
 Branched-chain amino acids cont.

20-Feb-25
53
 Phenylalanine and Tyrosine
• The degradation of phenylalanine and tyrosine are the same

– Phenylalanine is converted to tyrosine

– Tyrosine undergoes oxidative degradation.

• Degradation produces fumarate and acetoacetate.

• Deficiencies of different enzymes in the pathway result:

– phenylketonuria

– Tyrosinemia

– alcaptonuria.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 54

 Phenylalanine and Tyrosine cont.

Degradation of phenylalanine and tyrosine

20-Feb-25 By Kejela B.(B. Pharm, MSc) 55

 Phenylalanine and Tyrosine cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 56

 Tryptophan
• Tryptophan is oxidized to alanine, formate and acetyl CoA.

• Tryptophan is both glucogenic and ketogenic.

• NAD+ and NADP+ is produced from ring structure of

tryptophan.

• Therefore, tryptophan spares the dietary requirement for

niacin.

– The higher the dietary levels of tryptophan, the lower the

levels of niacin required

to prevent symptoms of deficiency.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 57
 Tryptophan cont.
• Tryptophan is both glucogenic and ketogenic.
• The indole ring is ketogenic and the side chain forms the
glucogenic product (alanine)
• kynurenic acid and xanthurenic acid are tryptophan-derived
isoquinolines
– not further degraded but are excreted in the urine.
– are in part responsible for the yellow color of urine

20-Feb-25 By Kejela B.(B. Pharm, MSc) 58

 Tryptophan cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 59

 Lysine

• Lysine is degraded to acetoacetyl-CoA in a pathway with

nine enzymatic reactions.

– it is ketogenic amino acid.

• Carnitine is synthesized from protein-bound trimethyllysine.

– transports long-chain fatty acids into the mitochondria

• After degradation of protein, trimethyllysine is converted to

carnitine
20-Feb-25 By Kejela B.(B. Pharm, MSc) 60
 Lysine cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 61

Intertissue amino acid metabolism and regulation

20-Feb-25 By Kejela B.(B. Pharm, MSc) 62

 Release of amino acids from skeletal muscle during fasting

• The efflux of amino acids from skeletal muscle supports the

essential amino acid pool in the blood

• Skeletal muscle oxidizes BCAAs (valine, leucine, isoleucine) to

produce energy and glutamine.

• The amino groups of the BCAAs, aspartate and glutamate

are transferred out of skeletal muscle in alanine and
glutamine.

• Alanine and glutamine account for approximately 50% of

the total α-amino nitrogen released by skeletal muscle.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 63

 Release of amino acids from skeletal muscle during fasting

• The release of amino acids from skeletal muscle is stimulated

by the decrease of insulin and increase of glucocorticoid
(cortisol) levels in the blood.
• Insulin promotes the uptake of amino acids and the general
synthesis of proteins.
• The fall of blood insulin levels results in net proteolysis and
release of amino acids.
• The release of glucocorticoid from the adrenal cortex
– induces ubiquitin synthesis
– increase of ubiquitin dependent proteolysis.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 64
 Release of amino acids from skeletal muscle during fasting

Interorgan amino acid exchange after an overnight fast.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 65
 Amino acid metabolism in liver during fasting
• The major site of alanine uptake is the liver
– disposes of the amino nitrogen by incorporating it into
urea
• The liver also extracts free amino acids, α-keto acids and
some glutamine from the blood.
• Alanine and other amino acids are oxidized and their carbon
skeletons converted principally to glucose.
• Glucagon and glucocorticoids stimulate the uptake of amino
acids into liver and increase gluconeogenesis and ureagenesis

20-Feb-25 By Kejela B.(B. Pharm, MSc) 66

 Amino acid metabolism in liver during fasting cont.
• Alanine transport into the liver is enhanced by glucagon.

• The induction of the synthesis of gluconeogenic enzymes by

glucagon and glucocorticoids related with:

– induction of many of the enzymes of amino acid

degradation

– induction of urea cycle enzymes

• Urea synthesis also increases because of the increased

supply of ammonium ions (NH4+)

– from amino acid degradation in the liver.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 67
 Amino acid metabolism in liver during fasting cont.

• Hormonal regulation of hepatic amino acid metabolism in

the postabsorptive state.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 68

 Metabolism of amino acids in other tissues during fasting

• Glucose produced by the liver is used for energy by the brain

and other glucose dependent tissues (erythrocytes).

• The muscle under conditions of exercise also oxidizes some of

this glucose to pyruvate

– which is used for the carbon skeleton of alanine.

– when AMP-activated protein kinase is active

• Glutamine is generated in skeletal muscle from the oxidation

of BCAAs

– for removal of NH4+ formed from amino acid catabolism

20-Feb-25 By Kejela B.(B. Pharm, MSc) 69
Metabolism of amino acids in other tissues during fasting

• The kidney, gut and cells with rapid turnover rates are the
major sites of glutamine uptake

• Glutamine serves as a fuel for these tissues

– serves as a nitrogen donor for purine synthesis

– serves as a substrate for ammoniagenesis in the kidney.

• Much of the unused nitrogen from glutamine is transferred to

pyruvate

– to form alanine in these tissues.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 70

Metabolism of amino acids in other tissues during fasting

• Alanine then carries the unused nitrogen back to the liver.

• The brain is glucose dependent

– but can also use BCAAs for energy.

• BCAAs also provide a source of nitrogen for neurotransmitter

synthesis during fasting.

• Other amino acids released from skeletal muscle protein

degradation also serve as precursors of neurotransmitters.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 71

Principles governing amino acid flux between tissues

20-Feb-25 By Kejela B.(B. Pharm, MSc) 72

 Principles governing amino acid flux between tissues

• The pattern of interorgan flux of amino acids is strongly

affected by:

– conditions that change the supply of fuels

the overnight fasting

a mixed meal, a high-protein meal

– conditions that increase the demand for amino acids

metabolic acidosis, surgical stress

traumatic injury, burns

wound healing, sepsis.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 73
Principles governing amino acid flux between tissues cont.
 Ammonia (NH3)
• Ammonia (NH3) is toxic.
• It is transported between tissues as alanine or glutamine.
• Alanine is the principal carrier of amino acid nitrogen
– from other tissues back to the liver
– nitrogen is converted to urea and excreted into the urine.
• The amount of urea synthesized is proportional to
the amount of amino acid carbon that is being
oxidized as a fuel.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 74

Principles governing amino acid flux between tissues cont.

• The differences in amino acid metabolism between tissues are

dictated by:

– the types and amounts of different enzyme

– the type and amount transport proteins present in each

tissue

– the ability of each tissue to respond to different

regulatory messages

hormones and neural signals

20-Feb-25 By Kejela B.(B. Pharm, MSc) 75

Principles governing amino acid flux between tissues cont.

• The pool of glutamine in the blood serves several essential

metabolic functions
• It provides ammonia for excretion of protons in the urine as
NH4+
• It serves as a fuel for the gut, kidney and cells of the immune
system.
• Glutamine is also required by the cells of the immune system
and other rapidly dividing cells
– in which its amide group serves as the source of nitrogen
for biosynthetic reactions.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 76
Principles governing amino acid flux between tissues cont.

• In the brain, formation of glutamine from glutamate and

NH4+ provides

– a means removing ammonia and transporting glutamate

between different cell types within the brain.
• During metabolic acidosis, kidney becomes the
predominant site of glutamine uptake
– at the expense of glutamine utilization in other tissues.
• During sepsis, cells involved in the immune response
(macrophages, hepatocytes) become the preferential sites of
glutamine uptake.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 77
Principles governing amino acid flux between tissues cont.

• The BCAAs form a significant portion of the composition of the

average protein

– can be converted to intermediates of tricarboxylic acid

(TCA) cycle

– used as fuels by almost all tissues.

• Valine and isoleucine are the major precursors of glutamine.

• The catabolism of amino acids occurs principally in the liver.

– Except for BCAAs, alanine, aspartate and glutamate

20-Feb-25 By Kejela B.(B. Pharm, MSc) 78

Principles governing amino acid flux between tissues cont.

• The ability to convert four-carbon intermediates of the TCA

cycle to pyruvate

– is required for oxidation of both BCAAs and glutamine.

– requires phosphoenolpyruvate (PEP) carboxykinase and,

– decarboxylating malate dehydrogenase

malic enzyme

– Most tissues have one or both of these enzymes.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 79

Principles governing amino acid flux between tissues cont.
• Amino acids are major gluconeogenic substrates

• Most of the energy obtained from their oxidation is derived

from oxidation of glucose formed from their carbon skeletons.

• A much smaller percentage of amino acid carbon is converted

to acetyl CoA or ketone bodies.

• The use of amino acids for synthesis of glucose for the brain
and other glucose-requiring tissues is subjected to:

– the hormonal regulatory mechanisms of glucose

homeostasis
20-Feb-25 By Kejela B.(B. Pharm, MSc) 80
Principles governing amino acid flux between tissues cont.

• The relative rates of protein synthesis and degradation

determine the size of the free amino acid pools

– which is available for the synthesis of new proteins and

for other essential functions.

• For example,

– synthesis of new proteins to mount an immune

response is supported by the net degradation of
other proteins in the body.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 81

 Utilization of amino acids in individual tissues

20-Feb-25 By Kejela B.(B. Pharm, MSc) 82

 Utilization of amino acids in individual tissues
A. Kidney
• One of the primary roles of amino acid nitrogen is to provide
ammonia in the kidney for the excretion of protons in the urine.
• NH4+ is released
– from glutamine by glutaminase
– from glutamate by glutamate dehydrogenase
both resulting in the formation of α-ketoglutarate.
• α-Ketoglutarate is used by kidney and oxidized to CO2.
– converted to glucose for use in the renal medulla
– converted to alanine to return ammonia to the liver
for urea synthesis
20-Feb-25 By Kejela B.(B. Pharm, MSc) 83
 Utilization of amino acids in individual tissues cont.
• Glutamine is used as a fuel by kidney in the normal fed state,
during fasting and metabolic acidosis.

• The carbon skeleton forms α-ketoglutarate

– which is oxidized to CO2, converted to glucose or released

as the carbon skeleton of serine or alanine.

• α-Ketoglutarate can be converted to oxaloacetate by TCA

cycle

• Oxaloacetate is converted to PEP by PEP carboxykinase.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 84

 Utilization of amino acids in individual tissues cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 85

 Utilization of amino acids in individual tissues cont.
• PEP is converted to pyruvate and acetyl CoA, alanine, serine
or glucose.
• The glucose is used by the cells of the renal medulla
– which have high dependence on anaerobic glycolysis
– because of their lower oxygen supply and mitochondrial
capacity.
• The lactate released from anaerobic glycolysis in these cells
is taken up and oxidized in the renal cortical cells
– which have higher mitochondrial capacity and greater
blood supply
20-Feb-25 By Kejela B.(B. Pharm, MSc) 86
 Utilization of amino acids in individual tissues cont.

Metabolism of glutamine and other fuels in the kidney

20-Feb-25 By Kejela B.(B. Pharm, MSc) 87
 Utilization of amino acids in individual tissues cont.
B. Skeletal Muscle
• Skeletal muscle is a major site of protein synthesis and
degradation.
• After high-protein meal, insulin promotes the uptake of certain
amino acids and stimulates net protein synthesis.
• The insulin stimulation of protein synthesis is dependent on an
adequate supply of amino acids to undergo protein synthesis.
• During fasting and other catabolic states
– there will be net degradation of skeletal muscle protein and
release of amino acids
20-Feb-25 By Kejela B.(B. Pharm, MSc) 88
 Utilization of amino acids in individual tissues cont.
• The net degradation of protein affects functional proteins
– which are sacrificed to meet more urgent demands for
amino acids in other tissues.
• During sepsis, degradation of skeletal muscle protein is
stimulated by the glucocorticoid (cortisol).
• The effect of cortisol is exerted through the activation of
ubiquitin-dependent proteolysis.
• During fasting, the decrease of blood insulin levels and the
increase of blood cortisol levels increase net protein
degradation
20-Feb-25 By Kejela B.(B. Pharm, MSc) 89
 Utilization of amino acids in individual tissues cont.
• Skeletal muscle is major site of glutamine synthesis thereby
satisfying the demand for glutamine
– during the postabsorptive state
– during metabolic acidosis
– during septic stress and trauma.
• The carbon skeleton and nitrogen of glutamine are derived
from metabolism of BCAAs.
• Amino acid degradation in skeletal muscle is also
accompanied by the formation of alanine
– which transfers amino groups from skeletal muscle to the
liver in the glucose-alanine cycle.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 90
 Utilization of amino acids in individual tissues cont.
 Oxidation of branched-chain amino acids in skeletal muscle

• The transamination of the BCAAs to α-keto acids occurs principally

in brain, heart, kidney and skeletal muscles.

• These tissues have high content of BCAAs transaminase relative to

low levels in liver.

• The α-keto acids of the BCAAs are then either

– released into the blood and taken up by liver OR,

– oxidized to CO2 or glutamine within the muscle or other tissue

• The oxidative pathways of the BCAAs convert the carbon

skeleton to either succinyl CoA or acetyl CoA
20-Feb-25 By Kejela B.(B. Pharm, MSc) 91
 Utilization of amino acids in individual tissues cont.
• The pathways generate NADH and FADH2 for ATP synthesis
before conversion of carbon to intermediates of the TCA cycle

– thus providing energy for muscle without loss of

carbon as CO2.
• Leucine is converted to acetyl CoA and acetoacetate.

• Skeletal muscle, adipocytes and most other tissues oxidize

leucine directly to CO2.

• The portion of isoleucine that is converted to acetyl CoA is

also oxidized directly to CO2.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 92
 Utilization of amino acids in individual tissues cont.
• For the portion of valine and isoleucine that enters the TCA
cycle as succinyl CoA to be completely oxidized to CO2

– it must first be converted to acetyl CoA.

• Succinyl CoA is oxidized to malate in the TCA cycle and

malate is then converted to pyruvate by malic enzyme

• Pyruvate can then be oxidized to acetyl CoA.

• Alternatively, pyruvate can form alanine or lactate.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 93

 Utilization of amino acids in individual tissues cont.

• Metabolism of the carbon

skeletons of BCAAs in skeletal

muscle.

• The oxidative pathways

generate NADH and FADH2

even before the carbon

skeleton enters the TCA cycle.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 94

 Utilization of amino acids in individual tissues cont.
 Conversion of branched-chain amino acids to glutamine
• The major route of valine and isoleucine catabolism in skeletal
muscle is to enter the TCA cycle as succinyl CoA
– exit as α-ketoglutarate to provide the carbon skeleton for
glutamine formation.
• Some of the glutamine and CO2 that are formed from net protein
degradation in skeletal muscle
– may also arise from the carbon skeletons of aspartate and
glutamate.
• These amino acids are transaminated and become part of the
four-carbon intermediates of the TCA cycle.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 95
 Utilization of amino acids in individual tissues cont.
• Glutamine nitrogen is derived principally from the BCAAs.

• The α-amino group released from transamination to form

glutamate from α-ketoglutarate

– the amide nitrogen is formed from the addition of free

ammonia to glutamate by glutamine synthetase.

• Free ammonia in the skeletal muscle is obtained from the

deamination of glutamate

– by glutamate dehydrogenase OR

– from the purine nucleotide cycle

20-Feb-25 By Kejela B.(B. Pharm, MSc) 96
 Utilization of amino acids in individual tissues cont.

Formation of glutamine from the amino groups of BCAAs

20-Feb-25 By Kejela B.(B. Pharm, MSc) 97

 Utilization of amino acids in individual tissues cont.
 Glucose-alanine cycle
• The nitrogen released from the oxidation of BCAAs in
skeletal muscle can be transferred back to the liver as
alanine
– in the glucose-alanine cycle
• Amino group of BCAAs is first transferred to α-ketoglutarate
to form glutamate
– then transferred to pyruvate to form alanine by sequential
transamination.
• The pyruvate is released from glucose via the glycolytic
pathway.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 98
 Utilization of amino acids in individual tissues cont.
• The alanine released from skeletal muscle is taken up by the
liver

– its amino group is incorporated into urea

– its carbon skeleton can be converted back to glucose

through gluconeogenesis.

• Although the amount of alanine formed varies with dietary

intake and physiological state

– transport of nitrogen from skeletal muscle to liver as alanine

occurs almost continuously throughout fasting-feeding cycle
20-Feb-25 By Kejela B.(B. Pharm, MSc) 99
 Utilization of amino acids in individual tissues cont.

Glucose-alanine cycle

20-Feb-25 By Kejela B.(B. Pharm, MSc) 100

 Utilization of amino acids in individual tissues cont.
C. Gut

• Amino acids are fuel for the intestinal mucosal cells after a
protein containing meal and in catabolic states

– during fasting or surgical trauma

• During fasting, glutamine is one of the major amino acids used

by the gut.

• The fates of glutamine carbon in the gut are oxidation to CO2

– then conversion to the carbon skeletons of lactate, citrulline,

and ornithine
20-Feb-25 By Kejela B.(B. Pharm, MSc) 101
 Utilization of amino acids in individual tissues cont.
• Nitrogen derived from amino acid degradation is converted to
citrulline, alanine and NH4+
• Although most of the carbon in alanine is derived from glucose
– oxidation of glucose to CO2 is not a major fuel pathway for the
gut.
• Fatty acids are also not a significant source of fuel for the intestinal
mucosal cells.
• After a protein meal, glutamine is a major fuel for the gut
• The gut also uses dietary aspartate and glutamate
• Colonocytes also use short-chain fatty acids derived from bacterial
action in the lumen.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 102
 Utilization of amino acids in individual tissues cont.

Amino acid metabolism in the gut

20-Feb-25 By Kejela B.(B. Pharm, MSc) 103

 Utilization of amino acids in individual tissues cont.
D. Liver

• The liver is the major site of amino acid metabolism.

• converts most of the carbon in amino acids

– to intermediates of the TCA cycle

– to acetyl CoA and ketone bodies.

• The liver is also the major site for urea synthesis.

• It can take up glutamine and alanine and convert the

nitrogen to urea for disposal.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 104

 Utilization of amino acids in individual tissues cont.
• The liver synthesizes plasma proteins

– serum albumin

– transferrin

– proteins of the blood coagulation cascade.

• Liver is a major site for:

– the synthesis of nonessential amino acids

– the conjugation of xenobiotic compounds with glycine

– the synthesis of heme and purine nucleotides

– the synthesis of glutathione.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 105
 Utilization of amino acids in individual tissues cont.
E. Brain and nervous tissue
• A major function of amino acid metabolism in neural tissue
is the synthesis of neurotransmitters
• These nurotransmitters are:
– amino acids (glutamate, glycine)
– catecholamines (dopamine, norepinephrine, epinephrine)
from tyrosine
– serotonin (from tryptophan)
– GABA (from glutamate)
– acetylcholine (from choline and acetyl CoA)
20-Feb-25 By Kejela B.(B. Pharm, MSc) 106
 Utilization of amino acids in individual tissues cont.
• The rapid metabolism of neurotransmitters requires the
continuous availability amino acids

– for de novo neurotransmitter synthesis.

• The brain is a net glutamine producer owing to the presence of

glutamine synthetase

• Glutamate and aspartate are synthesized in brain using

– amino groups donated by the BCAAs (valine)

– TCA cycle intermediates

– carbon skeletons of BCAAs

20-Feb-25 By Kejela B.(B. Pharm, MSc) 107
 Utilization of amino acids in individual tissues cont.

• The glutamate is converted to glutamine by glutamine

synthetase

• Glutamine synthase incorporates NH4+ released from:

– deamination of amino acids

– deamination of AMP in the purine nucleotide cycle in the

brain.

• This glutamine carries excess NH4+ from brain into blood OR,

– serve as a precursor of glutamate in neuronal cells

20-Feb-25 By Kejela B.(B. Pharm, MSc) 108

 Utilization of amino acids in individual tissues cont.
• Glutamine synthesized in brain is precursor
– for glutamate (excitatory neurotransmitter)
– for GABA (inhibitory neurotransmitter) in the neuronal cells
• In GABA-ergic neurons, glutamate is decarboxylated to GABA
– GABA is released during excitation of the neuron.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 109

 Disorders of amino acid metabolism

20-Feb-25 By Kejela B.(B. Pharm, MSc) 110

 Disorders of amino acid metabolism

20-Feb-25 By Kejela B.(B. Pharm, MSc) 111

 Disorders of amino acid metabolism cont.
 Homocystinemia/Homocystinuria

• Cystathionine synthase deficiency results in the accumulation

of homocysteine, homocystine and methionine

• Accumulation of homocystine in blood is associated with:

– cardiovascular disease

deep vein thrombosis

Thromboembolism, stroke

– dislocation of the lens (ectopic lens)

– mental retardation
20-Feb-25 By Kejela B.(B. Pharm, MSc) 112
 Disorders of amino acid metabolism cont.
• Homocystinuria similar to Marfan syndrome.

• Marfan syndrome is defect in the fibrillin gene and results:

– tall stature, long fingers and toes

– lens dislocation and tendency toward aortic wall

ruptures.

• In cystathionine synthase deficiency partial displacement of

the lens is downward and inward.

• In Marfan syndrome partial displacement of the lens is

upward and outward.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 113
 Disorders of amino acid metabolism cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 114

 Disorders of amino acid metabolism cont.
 Albinism

• Albinism is a group of conditions in which then normal

conversion of tyrosine to melanin is altered.

• The most severe form is deficiency of tyrosinase

• Results an absence of pigment in the skin, hair and eyes.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 115

 Disorders of amino acid metabolism cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 116

 Disorders of amino acid metabolism cont.
 Maple Syrup Urine Disease

• Caused by deficiency of branched-chain α-keto acid

dehydrogenase

– which oxidatively decarboxylates all three branched-

chain α-keto acids

• Characterized by elevation of plasma levels of:

– valine and its α-keto acids

– leucine and its α-keto acids

– isoleucine and its α-keto acids

20-Feb-25 By Kejela B.(B. Pharm, MSc) 117
 Disorders of amino acid metabolism cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 118

 Disorders of amino acid metabolism cont.
 Phenylketonuria (PKU)
• PKU is caused by complete deficiency of phenylalanine
hydroxylase.
• Phenylalanine is accumulated
– most of the accumulated phenylalanine is transaminated to
phenylpyruvate.
• Transamination is otherwise minor pathway of phenylalanine
metabolism.
• Phenylpyruvate is converted to other products that are
excreted in the urine along with phenylalanine and
phenylpyruvate
• Characterized by mental retardation and hypopigmentation
20-Feb-25 By Kejela B.(B. Pharm, MSc) 119
 Disorders of amino acid metabolism cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 120

 Disorders of amino acid metabolism cont.
 Alkaptonuria
• Alkaptonuria is inborn error of tyrosine degradation in which
accumulates homogentisate
• Homogentisate in the urine is oxidized and polymerized to
black products on exposure to light and air.
• Black pigment gradually accumulates in cartilage and other
connective tissues (ochronosis)
• Type-I tyrosinemia is caused by deficiency of
fumarylacetoacetate hydrolase
• Fumarylacetoacetate and related organic acids accumulate
20-Feb-25 By Kejela B.(B. Pharm, MSc) 121
 Disorders of amino acid metabolism cont.
• Tyrosinemia-I (tyrosinosis) is associated with liver failure,
cabbagelike odor and death.
• Tyrosinemia-II is caused by genetic deficiency of tyrosine
aminotransferase (TAT)
– lead to lesions of the eye and skin as well as
neurological problems
 Cystinuria
• caused by inherited defect in the intestinal absorption and
renal reabsorption of the dibasic amino acids
– lysine, arginine, ornithine and cystine.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 122
 Disorders of amino acid metabolism cont.
 Nonketotic hyperglycinemia

• It is recessive inherited disease

• It is caused by functional absence of the glycine-cleaving

enzyme

 Histidinemia

• It is recessive inherited condition in which histidase is deficient.

• Histidine levels in the blood are elevated and transamination

becomes histidine catabolism

20-Feb-25 By Kejela B.(B. Pharm, MSc) 123

 Disorders of amino acid metabolism cont.
 Methylmalonic aciduria

• It is caused by inherited defect in the vitamin B12-dependent

methylmalonyl-CoA mutase.

• Methylmalonic acid accumulates in the blood and is excreted

in the urine.

• leads to hypoglycemia, ketosis and hyperammonemia.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 124

 Disorders of amino acid metabolism cont.
 Hartnup Disease

• Hartnup disease is inherited defect in intestinal absorption

and renal reabsorption of large neutral amino acids

• Characterized by pellagera, dermatitis and neurological

abnormalities

• These signs are caused by decreased availability of

tryptophan

• Normally, niacin requirement is covered by endogenous

synthesis from tryptophan.

• Therefore tryptophan deficiency can cause niacin deficiency.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 125
 Disorders of amino acid metabolism cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 126

 Urea cycle

20-Feb-25 By Kejela B.(B. Pharm, MSc) 127

20-Feb-25 By Kejela B.(B. Pharm, MSc) 128
 Urea cycle
• Urea is the major nitrogenous excretory product which exits
from the body in the urine
• Urea is harmless compound produced mainly in the liver by
the urea cycle
– serves as the disposal form of ammonia which is toxic to
the brain and central nervous system.
• The level of ammonia in the blood is 30-60 μM.
• Ammonia is rapidly removed from the blood and converted
to urea by the liver.
• Nitrogen in the blood travels mainly by alanine and
glutamine.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 129
 Urea cycle cont.
• Nitrogen enters the urea cycle as NH4+ and aspartate.
• NH4+ forms carbamoyl phosphate which reacts with ornithine
to form citrulline.
• Ornithine is the compound that initiates and regenerated by
the cycle.
• Aspartate reacts with citrulline and donates nitrogen for urea
formation.
• Arginine is formed in two successive steps.
• Cleavage of arginine by arginase releases urea and
regenerates ornithine.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 130
20-Feb-25 By Kejela B.(B. Pharm, MSc) 131
20-Feb-25 By Kejela B.(B. Pharm, MSc) 132
 Steps in urea cycle
1. Synthesis of carbamoyl phosphate
• NH4+, HCO3- and ATP react to form carbamoyl phosphate.
• 2 ATP are hydrolyzed to form carbamoyl phosphate (high-
energy phosphate bond).
• Carbamoyl phosphate synthetase I (CPSI)
– found mainly in mitochondria of the liver and intestine
– produces carbamoyl phosphate for synthesis of urea
• Carbamoyl phosphate synthetase II (CPSII)
– located in the cytosol produces carbamoyl phosphate
– for pyrimidine biosynthesis using nitrogen from glutamine
20-Feb-25 By Kejela B.(B. Pharm, MSc) 133
 Steps in urea cycle cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 134

 Steps in urea cycle cont.
2. Production of arginine by the urea cycle
• Carbamoyl phosphate reacts with ornithine to form citrulline
• The high-energy phosphate bond of carbamoyl phosphate
provides the energy required for this reaction
– which occurs in mitochondria and is catalyzed by
ornithine transcarbamoylase (OTC).
• The citrulline is transported across the mitochondrial
membranes
– in exchange for cytoplasmic ornithine and enters the
cytosol.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 135
 Steps in urea cycle cont.
• The carrier for this transport reaction catalyzes an electro
neutral exchange of the two compounds.

• In the cytosol, citrulline reacts with aspartate to produce

argininosuccinate.

• Aspartate is the second source of nitrogen for urea

synthesis.

• Catalyzed by argininosuccinate synthetase

• Hydrolysis of ATP to AMP and pyrophosphate.

• Aspartate is produced by transamination of oxaloacetate.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 136
 Steps in urea cycle cont.
• Argininosuccinate is cleaved by argininosuccinate lyase to
form fumarate and arginine.
• Fumarate is produced from the carbons of argininosuccinate
provided by aspartate.
• Fumarate is converted to malate (by cytoplasmic fumarase)
– used for the synthesis of glucose or regeneration of
oxaloacetate by cytoplasmic reactions
• Oxaloacetate formed is transaminated to generate the
aspartate that carries nitrogen into the urea cycle.
• Thus, the carbons of fumarate can be recycled to aspartate.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 137
 Steps in urea cycle cont.

3. Cleavage of arginine to produce urea

• Arginine contains nitrogens derived from NH4 and aspartate

– cleaved by arginase to urea and ornithine.

• Urea is produced from the guanidinium group on the side

chain of arginine.

• The portion of arginine originally derived from ornithine is

reconverted to ornithine.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 138
 Steps in urea cycle cont.

• The conversion of citrulline to arginine and the cleavage of

arginine to urea occur in the cytosol.

• Ornithine is transported into the mitochondrion in exchange for

citrulline

– where it can react with carbamoyl phosphate

– initiates another round of the cycle.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 139

 Origin of ornithine

• Ornithine is an amino acid.

• However, it is not incorporated into proteins during the

process of protein synthesis

– because no genetic codon exists for this amino acid.

• Although ornithine is normally regenerated by the urea cycle

– ornithine also can be synthesized de novo from

glutamate semialdehyde

in an unusual transamination reaction in the intestine.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 140
 Regulation of Urea Cycle

20-Feb-25 By Kejela B.(B. Pharm, MSc) 141

 Regulation of the Urea Cycle
• The liver has a vast capacity to convert amino acid nitrogen
to urea
– thereby prevents toxic effects from ammonia
• Urea cycle is regulated by
– substrate availability
– higher the rate of ammonia production
– higher is the rate of urea formation.
• Regulation by substrate availability is a general characteristic
of disposal pathways.
– remove toxic compounds from the body.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 142
 Regulation of the urea cycle cont.
• In contrast to the feedback regulation, this is a type of feed-
forward regulation
– that produce functional end products.
• Two other types of regulation control the urea cycle.
– allosteric activation of CPSI

by N-acetylglutamate (NAG)

– induction and repression of the synthesis of urea cycle
enzymes.
• NAG is formed specifically to activate CPSI

20-Feb-25 By Kejela B.(B. Pharm, MSc) 143

 Regulation of the urea cycle cont.
• The synthesis of NAG from acetyl CoA and glutamate is
stimulated by arginine.

• Thus, two important reactions are stimulated as arginine

levels increase within the liver.

• The first is the synthesis of NAG

– increases the rate of carbamoyl phosphate production.

• The second is to produce more ornithine by arginase

– that can operate the cycle more rapidly.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 144

 Regulation of the urea cycle cont.

• The induction of urea cycle enzymes occurs in response to

conditions that require increased protein metabolism

– such as high-protein diet or prolonged fasting.

• In both states, as amino acid carbon is converted to glucose,

amino acid nitrogen is converted to urea.

• The ability of high-protein diet to increase urea cycle

enzyme levels is another type of feed-forward regulation

20-Feb-25 By Kejela B.(B. Pharm, MSc) 145
 Function of the urea cycle during fasting
• During fasting, the liver maintains blood glucose levels.
• Amino acids from muscle protein are major carbon source for
the production of glucose by gluconeogenesis.
• As amino acid carbons are converted to glucose, the
nitrogen are converted to urea.
• Thus, the urinary excretion of urea is high during fasting.
• As fasting progresses, the brain begins to use ketone bodies
and spares blood glucose.
• Less muscle protein is cleaved to provide amino acids for
gluconeogenesis
– decreases production of glucose and urea from amino acids
20-Feb-25 By Kejela B.(B. Pharm, MSc) 146
 Function of the urea cycle during fasting cont.
• The major amino acid substrate for gluconeogenesis is alanine
– synthesized in peripheral tissues to act as a nitrogen
carrier.
• Glucagon release stimulates alanine transport into the liver
– by activating the transcription of transport systems for
alanine.
• Two molecules of alanine are required to generate one
molecule of glucose.
• The nitrogen from the two molecules of alanine is converted
to one molecule of urea.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 147
 Disorders of the urea cycle

20-Feb-25 By Kejela B.(B. Pharm, MSc) 148

 Disorders of the urea cycle
• Disorders of the urea cycle are hazardous
– due to the accumulation of ammonia in the circulation.
• Ammonia is toxic to the nervous system
– its concentration in the body must be carefully controlled.
• Under normal conditions, free ammonia is rapidly fixed into
either:
– α-ketoglutarate to form glutamate
by glutamate dehydrogenase
– glutamate to form glutamine
by glutamine synthetase
20-Feb-25 By Kejela B.(B. Pharm, MSc) 149
 Disorders of the urea cycle cont.

• The glutamine can be used by many tissues

• Glutamate donates nitrogen to pyruvate to form alanine

which travels to the liver

• Within the liver, the nitrogen are removed from their carriers

– CPSI fixes the ammonia into carbamoyl phosphate to

initiate the urea cycle.

• When urea cycle enzyme is defective the cycle is interrupted

– leads to an accumulation of urea cycle intermediates.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 150

 Disorders of the urea cycle cont.
• Because of the block in the urea cycle:
– glutamine levels increase in the circulation
 due to α-ketoglutarate is no longer being regenerated by
removal of nitrogen from glutamine
 α-ketoglutarate levels are too low to fix more free
ammonia
leads to elevated ammonia levels in the blood.
• Therefore, defects in urea cycle enzyme lead to elevated
glutamine and ammonia levels.
• Extent of the elevation depends on which enzyme is defective
20-Feb-25 By Kejela B.(B. Pharm, MSc) 151
 Disorders of the urea cycle cont.
• Ammonia toxicity lead to brain swelling
– due to high levels of ammonia and glutamine in astrocytes.
• As ammonia levels increase in the astrocytes
– more glutamine is produced by glutamine synthetase
– which only exacerbates the osmotic imbalance.
• The ammonia levels inhibit glutaminase
– leading to glutamine elevation
• The high levels of glutamine alter the permeability of the
mitochondrial membrane
– leading to an opening of a pore in the mitochondrial
membrane which leads to cell death.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 152
 Disorders of the urea cycle cont.
• Another toxic effect of ammonia is lowering of glutamate
levels
– due to the high activity of the glutamine synthetase.
• Glutamate is neurotransmitter
• Impairment of glutamatergic neurotransmission causes brain
dysfunction because:
– glutamate is one of the excitatory neurotransmitters
– the absence of glutamate neurotransmission results lethargy
and reduced nervous system activity.
• The most common urea cycle defect is OTC deficiency and it is
an X-linked disorder.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 153
 Disorders of the urea cycle cont.
• The major clinical problem in treating patients with urea cycle
defects is reducing excessive blood ammonia on the nervous
system.
• High ammonia levels can lead to irreversible neuronal damage
and mental retardation.
• Low-protein diets are essential to reduce the potential for
excessive amino acid degradation.
• If the enzyme defect in the urea cycle comes after the synthesis
of argininosuccinate
– arginine supplementation proves the defect.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 154
 Disorders of the urea cycle cont.
• Once argininosuccinate has been synthesized
– two nitrogen molecules intended for excretion is
incorporated into the substrate
– but there is no regeneration of ornithine.
• If ornithine is replenished, argininosuccinate is used as the
carrier for nitrogen excretion from the body.
• Thus, ingesting large levels of arginine leads to ornithine
production and nitrogen excretion by the arginase
• Arginine therapy will not work for enzyme defects that exist
before the synthesis of argininosuccinate.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 155
 Disorders of the urea cycle cont.
• For these disorders, drugs that form conjugates with amino
acids are used .
• The conjugated amino acids are excreted and the body then
has to use its nitrogen to resynthesize the excreted amino acid.
• The two compounds most frequently used are benzoic acid and
phenylbutyrate.
• The active component of phenylbutyrate is phenylacetate, its
oxidation product.
• Phenylacetate has a bad odor
– this makes it difficult to take orally.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 156
 Disorders of the urea cycle cont.
• After activation, benzoic acid reacts with glycine to form
hippuric acid which is excreted.
• As glycine is synthesized from serine, the body now uses
nitrogens to synthesize serine
– so more glycine can be produced.
• Phenylacetate forms a conjugate with glutamine which is
excreted.
• This conjugate removes two nitrogens per molecule and
requires the body to resynthesize glutamine from glucose
– thereby using another two nitrogen molecules.
• Urea cycle defects are excellent candidates for treatment by
gene therapy.
20-Feb-25 By Kejela B.(B. Pharm, MSc) 157
 Disorders of the urea cycle cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 158

 Disorders of the urea cycle cont.

20-Feb-25 By Kejela B.(B. Pharm, MSc) 159

20-Feb-25 By Kejela B.(B. Pharm, MSc) 160