2.

05
July 29, 2016

CATABOLISM OF PROTEINS AND OF AMINO ACID NITROGEN

Raymond Oliver D. Cruz, MD, FPAFP

 Subsequent deamination of glutamine in the liver releases

TOPIC OUTLINE ammonia, which is then converted to nontoxic urea.
I. Biomedical Importance  If liver function is compromised, as in cirrhosis or hepatitis,
II. Protein turnover occurs in all life forms elevated blood ammonia levels generate clinical signs and
III. Proteases and peptidases degrade proteins to Amino Acids symptoms
a. Ubiquitin
IV. Ubiquitin-Proteasome mechanism
a. Glucose-Ala Cycle
b. Digestion of dietary protein
i. Stomach
ii. Pancreas
iii. Small Intestine
V. Animals convert Amino Nitrogen to Varied end products
VI. Transamination transfers
a. Transamination
b. Pyridoxal Phosphate
VII. Alanine pyruvate aminotransferase and glutamate
aminotransferase
VIII. L-glutamate dehydrogenase occupies a central position in
Nitrogen metabolism
IX. Amino Acid Oxidases Also Remove Nitrogen as Ammonia
a. Ammonia Intoxication Is Life- Threatening
X. Glutamine Synthase Fixes Ammonia as Glutamine
XI. Glutaminase & Asparaginase Deamidate Glutamine &
Asparagine
XII. Formation & secretion of Ammonia maintains Acid-base
balance
XIII. The Urea Cycle
a. Function
b. Reactions
c. Urea is the major end product of nitrogen catabolism
in humans  Whenever you take protein, it is digested in the stomach the
d. Condensation of CO2, ammonia, and ATP to form digestive enzyme is called your Pepsin which is only active at a
carbamoyl phosphate low pH. The reason why you have low pH on your stomach because
e. Formation of Argininosuccinate from Citrulline and these enzymes which digests your proteins needs to be activated.
Aspartate These enzymes, at a low pH will release your amino acid.
f. Formation of Arginine and Fumarate from cleavage  Most of these enzymes are secreted at gastrointestinal tract
of Argininosuccinate specifically the pancreas so these pancreatic enzymes will release
XIV. Argininoosuccinae smaller segments of the proteins until they reach the small intestine
XV. Cleavage of Arginase Releases Urea and re-forms Ornithine and then your intestinal di and tri peptidases will try to break it
XVI. Arginase down to your amino acid.
XVII. Carbamoyl Phosphate synthase I is the pacemaker enzyme  In small intestine there are a different digestive enzymes and in the
XVIII. N-acetylglutamate synthase initial digestive that is usually activated is Trypsin because it
XIX. Metabolic Disorders activates all of the other digestive enzymes.
XX. Gene therapy offers promise for correcting defects in urea o Remember: What activates trypsin? The precursor of trypsin is
biosynthesis trypsinogen and this is what you call a zymogen.

BIOMEDICAL IMPORTANCE
 Protein and amino acid catabolism describes how the nitrogen of
amino acids is converted to urea
 In normal adults, nitrogen intake matches nitrogen excreted
 In diseased conditions, this balance is affected
 Positive nitrogen balance, an excess of ingested over excreted
nitrogen, accompanies growth and pregnancy.
 Negative nitrogen balance, where output exceeds intake, may
follow surgery, advanced cancer, and kwashiorkor or marasmus
 While ammonia, derived mainly from the á-amino nitrogen of
amino acids, is highly toxic, tissues convert ammonia to the amide
nitrogen of nontoxic glutamine.

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 2.05
TITLE OF LECTURE (NOTE: ALL CAPS)
TRYPSIN
 Digestive enzyme that breaks down protein into smaller products
 Intial digestive enzyme that activates all of zymogen (an inactive
substance that is converted into an enzyme when activated by
another enzyme) such as Trypsonogen, Chymotrypsinogen,
Proelastrate, and Procarboxypeptidase.
 Its precursor is TRYPSINOGEN, a zymogen, that is converted into
Trypepsin by ENTEROPEPSIDASE (an enzyme produced by the
gastrointestinal tract)
NOTE: “Trypsin has a specific area to cleave in the protein. If there is
already a basic protein present, it will cleave its carboxyl terinal Argenin
[Arg] and Lysin [Lys]” – Doc
CHYMOTRYPSIN
 Digestive enzyme that breaks down protein in to smaller products
 Its precursor is CHYMOTRYPSINOGEN
NOTE: “Chymotrypsin cleave its carboxyl terminal Trp, Tyr, Phe, Met, Leu,
and is for larger neutral amino acids” – Doc
ELASTASE
 Digestive enzyme that breaks down proteins into smaller products
 Its precursor is PROELASTASE
NOTE: “Elastase cleave its carboxyl terminal Ala, Gly, Ser and is for smaller
amino acids” - Doc
CARBOXYPEPTIDASES (A and B)
 Are digestive enzymes that break down proteins into smaller
products
 Their precursors are PROCARBOXYPEPTIDASES A and B,
respectively
NOTE: “Carboxypeptidase A cleaves its carboxy terminals Ala, Ile, Leu and
Val while Carboxypeptidase B cleaves its carboxy terminals Arg and Lyse.
Both are for last amino acids” - Doc
PROTEIN TURNOVER OCCURS IN ALL FORMS OF LIFE
 Each day, human’s turnover 1-2% of total body protein
NOTE:
“Excess proteins will be converted into fats” - Doc
“Excess proteins can cause a hard time for kidneys and may result to kidney
failure” - Doc
 High protein degradation occur in tissues undergoing structural
rearrangement such as uterine tissue during pregnancy, tadpole tail
tissue during metamorphosis, or skeletal muscle starvation
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Catabolism of Proteins and of Amino Acid Nitrogen
NOTE: “Pregnant women need positive protein intake for child growing” –
Doc
 Only 75% of liberated amino acids are reutilized
NOTE: “Old or worn out proteins are will be recycled, the 75%” – Doc
 The excess amino acids (15%) will be converted to nitrogen in for of
urea
 Excess amino acids are not stored, those are not immediately
incorporated into new proteins are rapidly degraded.
NOTE:
 “Amino acids are not stored, instead they are converted into
carbohydrates (sugars) which can be stored in the body”
 “We have no mechanism to store amino acid, except of it will be
incorporated into proteins” - Doc
 “Only a few specific situations such as the destruction of muscle then
the amino acids will be needed by the body” - Doc
PROTEASE AND PEPTIDASES DEGRADE PROTEINS TO
AMINO ACIDS
 The susceptibility of a protein to degradation is expressed as its
half-life (“the time within which the ½ of the concentration of the
protein has already been degraded”)
NOTE: “Drugs’ half-life is the basis of drug consumption such as: twice-a-
day, every 3hrs/8hrs/12hrs or so, and etc” – Doc
 Half-lives of proteins ranges from under .5hr (30mins) to over
150hrs
 Typically “housekeeping” enzymes (“enzymes that are maintained
or regularly produced) have shorter half-life values of just over
100hrs
 For enzymes that are need to be controlled such as rate-limiting
enzymes and enzymes that control key regulatory processes, then
the half-life becomes shorter from .5hr (30 ins) to 2hrs.
 PEST sequences, region of rich;
P- Proline
E- Glutamate
S- Serin
T- Theonine
target some proteins to peptide rapid degradation. The resulting
peptides are then degraded to amino acids by ENDOPEPTIDASE that
cleave internal bonds.
NOTE: “Some amino acids will be recycled, others will be degraded” - Doc
AMINOPEPTIDASES
 Remove amino acids sequentially from amino terminals
CARBOPEPTIDASES
 Remove amino acids sequentially from carboxyl terminals
 Extracellular, membrane-associated, and long-lived intracellular
proteins are degraded in Lysosomes by ATP-independent process
NOTE:
 “Long-lived Intracellular Proteins are your “housekeeping” proteins
and enzymes” - Doc
 “By contrast, degradation of abnormal and other short-lived proteins
occurs in the cytosol and requires ATP and Ubiquitin” - Doc
 2.05
A. Ubiquitin
 Ubiquitin is a small (8.5 kDa) protein that targets many intracellular
TITLE OF LECTURE (NOTE: ALL CAPS)
proteins for degradation.
NOTE: When proteins are said to be ubiquitinated, it means that the
proteins are ready for degradation.
 The primary structure of ubiquitin is highly conserved. Only 3 of 76
residues differ between yeast and human ubiquitin.
 Several molecules of ubiquitin are attached by non-peptide bonds
formed between the carboxyl terminal of ubiquitin and the amino
groups of lysyl residues in the target protein.
NOTE: End point of the 3 enzymes: to ubiquitinate protein so that it
will be degraded. More ubiquitin molecules are attached, the faster
will be the targeting.
 The terminal COOH of ubiquitin first forms a thioester
 The coupled hydrolysis of PPi by pyrophosphatase ensures that the
reaction will proceed readily
 A thioester exchange reaction transfers activated ubiquitin to E2.
 E3 then catalyzes the transfer of ubiquitin to the -amino group of a
lysyl residue of the target protein.
 Additional rounds of ubiquitination result in subsequent
polyubiquitination.
NOTE: Doc will not include the specific reactions 
UBIQUITIN-PROTEASOME MECHANISM
 The residue present at its amino terminal affects whether a protein
is ubiquitinated.
 Amino terminal Met or Ser retards whereas Asp or Arg accelerates
ubiquitination.
 Degradation occurs in an autocatalytic complex of proteases known
as the proteasome.
A. Glucose-Ala Cycle
NOTE: One of the most important amino acid in the process of converting
amino acid to sugar is alanine. Alanine is converted to pyruvate by the
removal of amino group. Lagi silang magkapartner because if you remove
the amino group of alanine it becomes pyruvate.
 The rate of hepatic gluconeogenesis from alanine is far higher than
from all other amino acids. *Fast because it’s a very easy process.
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Catabolism of Proteins and of Amino Acid Nitrogen
 The capacity of the liver for gluconeogenesis from alanine does not
reach saturation until the alanine concentration reaches 20–30
times its physiologic level.
 Following a protein-rich meal, the splanchnic tissues release amino
acids while the peripheral muscles extract amino acids, in both
instances predominantly branched- chain amino acids.
 Branched-chain amino acids serve a special role in nitrogen
metabolism, both in the fasting state, when they provide the brain
with an energy source, and after feeding, when they are extracted
predominantly by muscles
NOTE: Branched chain AA are preferentially absorbed by the muscles
especially after exercise.
A. Digestion of Dietary Proteins
 Proteins are hydrolyzed to amino acids for absorption to make them
smaller
 Proteolytic enzymes responsible for degrading proteins are in the
stomach, pancreas, and small intestine
STOMACH
 Contains gastric juices HCL and pepsin
 HCl kills bacteria
 Pepsin is from pepsinogen which is activated by HCl will digest
peptide linkages
PANCREAS
 Pancreatic proteases further cleave polypeptides initially digested
by the stomach after the contents of what you eat pas through your
duodenum and intestine.
 Enteropeptidase from the intestine activates pancreatic enzymes
such as trypsinogen to its active form.
SMALL INTESTINE
 Contains aminopeptidase that cleaves oligopeptides to produce free
amino acids
 Free amino acids and dipeptides are absorbed by the intestinal
epithelium
 Dipeptides are further hydrolyzed in the cytosol before being
released to the portal system
ANIMALS CONVERT AMINO NITROGEN TO VARIED END
PRODUCTS
 The aqueous environment of teleostean fish, which are ammonotelic
(excrete ammonia) (DIFFUSES THROUGH THE SKIN), compels them
to excrete water continuously, facilitating excretion of highly toxic
ammonia.
 Birds, which must conserve water and maintain low weight, are
uricotelic and excrete uric acid as semisolid guano.
 Many land animals, including humans, are ureotelic and excrete
nontoxic, water-soluble urea.
 High blood urea levels in renal disease are a consequence—not a
cause—of impaired renal function.
 Remember: LIVER synthesizes Urea and not the kidney!
Urea biosynthesis occurs in 4 stages:
1. Transamination - amino group is removed and becomes a
carbohydrate. Examples of transaminases are SGOT, SGPT, ALT, AST
2. Oxidative deamination of glutamate
NOTE: Glutamate is a transition amino acid because if you remove
glutamine amino group, it will become your alpha ketoglutarate which is
part of the Kreb’s cycle.
 Glutarate to Alpha ketoglutamate: important step in the
conversion of Urea
 2.05 Catabolism of Proteins and of Amino Acid Nitrogen

o These 2 enzymes catalyze the transfer of amino groups to

3. Ammonia transport - release ammonia and it is transported in the pyruvate (forming alanine) or to á - ketoglutarate (forming
TITLE
liver where you have OF LECTURE
the synthesis of ammonia. (NOTE: ALL CAPS) glutamate)
 Ammonia + CO2 = for the synthesis of Urea

4. Reactions of the urea cycles

 Remember: Amino Acid - Alpha keto acid partners

TRANSAMINATION TRANFERS
A. Transamination
 interconverts pairs of alpha amino acids and keto acids.
 All of the protein amino acids EXCEPT glycine, threonine, proline and
hydroxyproline participate in this transamination process.
 The reaction is reversible; pwedeng addition of amino group,
pwedeng tatanggalin naman yun amino group.
 SGPT, SGOT, AST, ALT – those are actually being transaminases.
NOTE: Pinapasa lang niya yung amino group from one substrate to
another. There is a need for a co-enzyme or a co-factor which is your
pyridoxal phosphate.
o Pyruvate – Alanine
 If you add an amino group to pyruvate it becomes
Alanine.
o Alpha ketoglutarate – Glutamate
o Oxaloacetate and aspartic acid
 Each aminotransferase is specific for one pair of substrates but
nonspecific for the other pair.
 Since alanine is also a substrate for glutamate aminotransferase, all
the amino nitrogen from amino acids that undergo transamination
can be concentrated in glutamate.
 This is important because L-glutamate is the only amino acid that
undergoes oxidative deamination at an appreciable rate in
mammalian tissues.
 The formation of ammonia from á -amino groups thus occurs
mainly via the á -amino nitrogen of L-glutamate
NOTE: Glutamate - the only amino acid that undergoes oxidative
deamination
 The alpha ketoglutarate will absorb or will condense with the
ammonia to be converted to glutamate.
o Or pwede pang madagdagan ng isa pang amino group yung
glutamate to become glutamine. And that glutamine can be
brought to the liver, release its ammonia and ammonia will be
converted to urea. Kaya very central ang function ng glutamate.

L-GLUTAMATE DEHYDROGENASE OCCUPIES A

CENTRAL POSITION IN NITROGEN METABOLISM
 Transfer of amino nitrogen to á -ketoglutarate forms L-
glutamate.
 Release of this nitrogen as ammonia is then catalyzed by hepatic L-
glutamate dehydrogenase (GDH), which can use either NAD+ or
NADP+

B. Pyridoxal Phosphate
 The coenzyme pyridoxal phosphate (PLP) is present at the catalytic
site of aminotransferases and of many other enzymes that act on
amino acids.
 During transamination, bound PLP serves as a carrier of amino
groups.
 Following removal of á -amino nitrogen by transamination, the
remaining carbon "skeleton" is degraded.
 Alanine-pyruvate aminotransferase (alanine aminotransferase) and
glutamate-alphaketoglutarate aminotransferase (glutamate
aminotransferase)

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NOTE: So the formation of ammonia from the alpha amino groups does
overstatedly via the alpha amino nitrogen of L-Glutamate
 The enzyme for thisTITLE OF LECTURE
process is your (NOTE: ALL CAPS) energy dahil kahit naka-upo ka or tulog ka laging gumagana ang brain.
L-Glutamate dehydrogenase.
o It uses NAD or NADP as absorber of your hydrogen. So NADP or
NAD acts as an oxidizing agent.
 Conversion of á -amino nitrogen to ammonia by the concerted
action of glutamate aminotransferase and GDH is often termed
"transdeamination."
 Liver GDH activity is allosterically inhibited by ATP, GTP, and NADH
and activated by ADP.
 The reaction catalyzed by GDH is freely reversible and functions also
in amino acid biosynthesis
NOTE: GDH will be active to deamination in converting amino acid to
glucose.
 Why? Kasi kulang ka sa energy. Diba glucose is a source of energy.
 Kung marami kang energy, marami kang ATP, GDP, NADH ganiyan,
you will need GDH. So di mo kailangan ng energy source. You don’t
need to convert our amino acids to glucose for energy. That’s why it
will be inihibted.
 However, when the cell lacks energy, it will activate now your GDH.
AMINO ACID OXIDASES ALSO REMOVE NITROGEN AS
AMMONIA
 L-amino acid oxidases of liver and kidney convert amino acids to an
á-imino acid that decomposes to an á-keto acid with release of
ammonium ion.
 The reduced flavin is reoxidized by molecular oxygen, forming
hydrogen peroxide, which then is split to O2 and H 2O by catalase.
NOTE: Catalase will breakdown hydrogen peroxide to water and it is
present in the peroxisomes.
A. Ammonia Intoxication is also Life-Threatening
 The ammonia produced by enteric bacteria and absorbed into
portal venous blood and the ammonia produced by tissues are
rapidly removed from circulation by the liver and converted to
urea.
 Thus, only traces (10–20 g/dL) normally are present in peripheral
blood.
 This is essential, since ammonia is toxic to the central nervous
system. It can cause Hyperammonemia – seizures, comatose, etc.
 Should portal blood bypass the liver, systemic blood ammonia
levels may rise to toxic levels. This occurs in severely impaired
hepatic function.
 Symptoms of ammonia intoxication include tremor, slurred
speech, blurred vision, coma, and ultimately death.
 Ammonia may be toxic to the brain in part because it reacts with α-
ketoglutarate to form glutamate.
 The resulting depleted levels of α-ketoglutarate then impair
function of the tricarboxylic acid (TCA) cycle in neurons.
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Catabolism of Proteins and of Amino Acid Nitrogen
NOTE: Uubusin yung α-ketoglutarate and the brain always needs that
Kapag naubusan siya ng α-ketoglutarate wala na siyang mechanism for the
the synthesis of energy, kasi lahat na convert na to glutamine. So, that will
explain the reason why it’s very toxic to brain tissue.
 Bakit ammonia ang ipinapa-amoy kapag nahihilo?
o Napro-process naman yun, para ma-stimulate lang yung nasal
passages during the inhalation of ammonia but it’s not
scientific.
o The irritation will lessen the symptoms of your dizziness or
headache kaya yun ang binibigay sa clinic pero wala ng ibang
reason para doon. Parang pang stimulate lang ng nasal
passages and hopefully will relieve the symptoms.
GLUTAMINE SYNTHASE FIXES AMMONIA AS
GLUTAMINE
 Formation of glutamine is catalyzed by mitochondrial glutamine
synthase.
 Since amide bond synthesis is coupled to the hydrolysis of ATP to
ADP and Pi, the reaction strongly favors glutamine synthesis.
 Glutamine synthase will synthesize your glutamine by
incorporating the ammonia into the structure of your glutamine.
So, this process is energy requiring.
 One function of glutamine is to sequester ammonia in a nontoxic
form.
NOTE:
 Why is Magnesium ATP complex always seen?
o The reason is because ATP is stabilized by Magnesium.
o Remember yung ATP madami siya phosphates na negatively
charged and Magnesium is positive kasi kung hindi maghihiwa-
hiwalay yung mga phosphates so useless yung ATP. So, that’s
why you will see Magnesium as a supplement in most Vitamins
that are for energy utilization (fortified with magnesium) so, yun
yung explanation non. The importance of Magnesium.
GLUTAMINASE & ASPARAGINASE DEAMIDATE
GLUTAMINE & ASPARAGINE
 Hydrolytic release of the amide nitrogen of glutamine as ammonia,
catalyzed by glutaminase strongly favors glutamate formation.
 The concerted action of glutamine synthase and glutaminase thus
catalyzes the interconversion of free ammonium ion and
glutamine.
 An analogous reaction is catalyzed by L-asparaginase
 2.05
TITLE OF LECTURE (NOTE: ALL CAPS)
NOTE: Kapag tinanggal ang ammonia, babalik uli yung glutamine to
glutamate just by the removals of amino group.
FORMATION & SECRETION OF AMMONIA MAINTAINS
ACID-BASE BALANCE
 Excretion into urine of ammonia produced by renal tubular cells
facilitates cation conservation and regulation of acid-base balance.
 Ammonia production from intracellular renal amino acids,
especially glutamine, increases in metabolic acidosis and
decreases in metabolic alkalosis.
NOTE:
 Kaya may effect yung protein intake with the acidity or alkalinity of
your urine. Minsan doon nadedetermine yung masyado kang
maraming tinake na protein so pwedeng ma-detect yun in the urine.
 Now, sometimes your urine is acidified in order to facilitate the
excretion of certain toxic chemicals for example, (walang aamin
siguro baka mabaril yung mga nag-shabu. Usong uso yan.) pupunta
sa ER kasi dilat ang pupils, walang tulog, mabilis ang heart rate.
Madaling namang ma-detect kung talagang intoxicated siya with
shabu/drugs o hindi. So, para ma-excrete yung metamphetamine
ina-acidify yung urine, you give Vitamin C. Pina-facilitate yun, in
order to remove the excess. Pero kung talagang nangdadaya ka sa
sports kunyari, kahit inom ka ng inom ng Vitamins C mahirap ma-
mask yun sa urine, mahuhuli at mahuhuli ka. So yung mahihilig dyan
sa fun run para lumakas titira ng shabu the night before para
walang tulugan, baka ma-heart attack kayo (haha).
THE UREA CYCLE (ORNITHINE CYCLE)
 The Urea Cycle is a series of reactions that produces urea from
ammonia (NH 3)
 This cycle was the first metabolic cycle discovered (Krebs and Kurt
Henseleit, 1932)
 In mammals, the urea cycle takes place only in the liver
A. The Function of Urea Cycle
 Organisms that cannot easily and quickly remove ammonia usually
have to convert it to some other substance, like urea or uric acid,
which are much less toxic
 Insufficiency of the urea cycle occurs in some genetic disorders
(inborn errors of metabolism), and in liver failure
 The result of liver failure is accumulation of nitrogenous waste,
mainly ammonia, which leads to hepatic encephalopathy (a
neuropsychiatric abnormality)
NOTE:
 Kapag nagkaron ka ng encephalopathy, ito yung kapag nakakakitaka
ng lasing nawawala sa sarili at nagkakaroon ng parang psychiatric
disorder, so sobrang kalasingan na di na nila alam yung ginagawa
nila.
 Kapag nakakita kayo ng liver cross-section ang daming mitochondria
kasi kailangan ng maraming energy and because of the importance of
urea cycle
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B. Reactions
 The urea cycle consists of five reactions – two mitochondrial and
three cytosolic
 The cycle converts two amino groups, one from NH 4+ and one from
Asp, and a carbon atom from HCO3-, to relatively nontoxic excretion
product, urea at the cost of four “high-energy" phosphate bonds
(3 ATP hydrolyzed to 2 ADP and 1 AMP)
 Ornithine is the carrier of these carbon and nitrogen atoms
NOTE:
 So, may reaction dito na conversion of your ADP to AMP at once. Ibig
sabihin ‘energy requirement is high’.
 So sa exam tandaan niyo kailangan niya ng 3 ATP para mag
synthesize pero ilang high energy phosphates ang ginagamit?
APAT/FOUR.
 Ito yung sinasabi natin kasi 3 ATP lang pero yung isang ATP is
converted to AMP and that will be 2 high energy phosphates at once.
 So, you need 3 ATPs but you need 4 high energy phosphates for the
process
C. Urea is the Major End Product of Nitrogen Catabolism in Humans
 Synthesis of 1 mol of urea requires 3 mol of ATP plus 1 mol each of
ammonium ion and of the α -amino nitrogen of aspartate
 N-acetylglutamate functions as an enzyme activator.
D. Condensation of CO2, ammonia, and ATP to form carbamoyl
phosphate
 catalyzed by: carbamoyl phosphate synthase I (CPS1)
o this is the rate-limiting enzyme of the urea cycle
o active only in the presence of its allosteric activator N-
acetylglutamate which enhances its affinity for ATP
o Defects in this enzyme are responsible "hyperammonemia
type 1" (rare metabolic disease)
 location: mitochondrial matrix (liver)
 requires 2 mol of ATP
D. Formation of Citrulline from Carbamoyl Phosphate and Ornithine
 transfer of the carbamoyl group of carbamoyl phosphate to
ornithine, forming citrulline and orthophosphate
 catalyzed by: L-Ornithine transcarbamoylase
o X-chromosome linked deficiency of this enzyme is termed
“hyperammonemia type 2"
 mothers exhibit hyperammonemia and an aversion to
high-protein foods
 elevated glutamine in blood, CSF, and urine, probably as a
result of enhanced glutamine synthesis in response to
elevated levels of tissue ammonia
 location: mitochondrial matrix (liver)
 Citrulline will then go to the cytoplasm
NOTE: Deficiency in Ornithine Transporter (due to mutation of the
ORNT1 gene) may lead to hyperornithinemia, hyperammonemia, and
homocitrullinuria syndrome (HHH syndrome). Failure to import cytosolic
ornithine into the mitochondrial matrix renders the urea cycle inoperable.
 Important: Abnormalities in ANY of the enzymes involved in the 5
reactions of Urea Cycle will lead to hyperammonemia. However,
deficiency in the enzymes from the first two reactions will cause
more severe hyperammonemia because these enzymes are
important in the degradation of ammonia.
 2.05
D. Formation of Argininosuccinate from Citrulline and Aspartate
 catalyzed by: Argininosuccinate synthase

TITLE OF LECTURE (NOTE: ALL CAPS)
aspartate is linked to citrulline via the amino group of aspartate--
provides the second nitrogen of urea (the other one is from the
free ammonia)
 location: liver cytosol
 requires ATP
o subsequent displacement of AMP by aspartate forms citrulline
E. Formation of Arginine & Fumarate from Cleavage of
Argininosuccinate
 catalyzed by: argininosuccinase
 location: liver cytosol
 proceeds with retention of nitrogen in arginine and release of the
aspartate skeleton as fumarate
o addition of water to fumarate forms L- malate, and
subsequent NAD+-dependent oxidation of malate forms
oxaloacetate (analogous to Kreb’s Cycle but are instead
catalyzed by cytosolic fumarase and malate dehydrogenase)
o transamination of oxaloacetate by glutamate aminotransferase
then re-forms aspartate
o aspartate will then be recycled to form more substrates
ARGININOSUCCINASE (ARGININOSUCCINATE LYASE)
 Arginine is produced from argininosuccinate
 The ASL gene is located on chromosome 7
 When deficient results in argininosuccinicaciduria, accompanied
by elevated levels of argininosuccinate in blood, cerebrospinal
fluid, and urine
 Argininosuccinicaciduria is associated with friable, tufted hair
(trichorrhexis nodosa)
(“Hindi ko to itatanong, hindi ko to matatandaan hindi ako interested”)
(Buti pa si Doc, nagsasabi ng hindi siya interested. Hindi siya paasa.
Tularan si Doc )
CLEAVAGE OF ARGININE RELEASES UREA AND RE-
FORMS ORNITHINE
 Hydrolytic cleavage of the guanidine group of arginine, catalyzed by
liver arginase, releases urea
 The other product, ornithine, reenters liver mitochondria for
additional rounds of urea synthesis.
 Ornithine and lysine are potent inhibitors of arginase, competitive
with arginine.
 Arginine also serves as the precursor of the potent muscle relaxant
nitric oxide (NO) in a Ca 2+ -dependent reaction catalyzed by NO
synthase
(“When you cleave off at least portions of your Arginine, you will now
release your Urea and form your Ornithine. Yung Orinithine papasok ulit to
the Ornithine transporter into the mitochondria in the liver para siya mag
condense with the carbomoyl phosphate to complete the cycle.”)
ARGINASE
 Hyperargininemia is an autosomal recessive defect in the gene for
arginase
 Unlike other urea cycle disorders, the first symptoms of
hyperargininemia typically do not appear until age 2 to 4 years.
o Blood and cerebrospinal fluid levels of Arginine are elevated
7 of 8 Catabolism of Proteins and of Amino Acid Nitrogen [Biöky3m3: Deiparine, Mendoza, Paat, Rocabo]
Catabolism of Proteins and of Amino Acid Nitrogen
(“Your arginase again cleaves ariginine releasing carbon and O nitrogens in
the form of urea so naform na yung urea to complete the cycle this is
referred to as hydrolysis wherein water is used to cleave
Your arginine is also a precursor for the potent muscle relaxant nitric oxide
Nitric oxide is important because it causes vasodilation of arteries and it
prevents onsent of hypertension.”)
CARBAMOYL PHOSPHATE SYNTHASE I IS THE
PACEMAKER ENZYME OF THE UREA CYCLE
 The activity of carbamoyl phosphate synthase I is determined by N-
acetylglutamate
 N-acetylglutamate level is dictated by its rate of synthesis from
Acetyl-CoA and glutamate and its rate of hydrolysis to acetate and
glutamate.
 These reactions are catalyzed by N- acetylglutamate synthase and
N- acetylglutamate hydrolase
(“Ayan sabi ni Doc, satsat daw to. Basahin nalang daw sa libro since hindi
naman importante. Ineemphasize lang daw niya yung mga importanteng
bagay)
(Buti pa si Doc, nakafocus lang sa kung ano yung importante. Marunong
mag bigay ng halaga. Tularan ulit si Doc )
N-ACETYLGLUTAMATE SYNTHASE
 N-Acetylglutamate is essential for carbamoyl phosphate synthase I
activity
 The NAGS gene encodes N-acetylglutamate synthase, which
catalyzes the condensation of Acetyl-CoA with glutamate.
 Defects in the NAGS gene result in severe hyperammonemia, which
may respond to administered N-acetylglutamate.
 Major changes in diet can increase the concentrations of individual
urea cycle enzymes 10- to 20-fold.
 Starvation elevates enzyme levels to cope with the increased
production of ammonia
(“N-acetyl glutamate NAGS gene – hyperammonemia kasi maaapektuhan
yung urea cycle
If you change your diet into a high protein diet the activity of the urea cycle
also increases because of the presence of protein so tumataas din yung urea
and tataas rin yung BUN tataas kasi mataas ang protein intake
BUN is more or less dependent on diet (also tests kidney fxn although it is
not as accurate as creatinine kasi creatinine dependent on muscle mass
kaya mas constant pero parehas sila kidney function)
Starvation elevates enzyme levels para tumaas rin levels ng ammonia
So during starvation, tataas rin levels ng urea.
Pag first step palang di na naincorporate ammonia mas maraming
ammonia mag aaccumulate so symptoms become more severe”)
METABOLIC DISORDERS OF THE UREA CYCLE
 Clinical symptoms common to all urea cycle disorders include
vomiting, avoidance of high-protein foods, intermittent ataxia,
irritability, lethargy, and severe mental retardation.
 Significant improvement and minimization of brain damage
accompanies a low-protein diet ingested as frequent small meals to
avoid sudden increases in blood ammonia levels.
 2.05 Catabolism of Proteins and of Amino Acid Nitrogen

GENE THERAPY OFFERS PROMISE FOR CORRECTING

DEFECTS IN UREA
TITLE OFBIOSYNTHESIS
LECTURE (NOTE: ALL CAPS)
 Gene therapy for rectification of defects in the enzymes of the urea
cycle is an area of active investigation.
 Encouraging preliminary results have been obtained, for example, in
animal models using an adenoviral vector to treat citrullinemia

(“Most of the defects are gene – gene therapy offers a lot of promise
Whisper Mechanism – Tinatanggal affected gene and replaced by the
connect genetic sequence”)

REFERENCES
 Dr. Cruz’ PPT and lecture

TRANSER’S MESSAGE

8 of 8 Catabolism of Proteins and of Amino Acid Nitrogen [Biöky3m3: Deiparine, Mendoza, Paat, Rocabo]