Interconversion of GLN, GLU, and -KG are important in

the regulation of blood NH3 concentration.

Ammonia is highly toxic to the CNS.Its level in the blood should be low.

Enzymes involved:
Transaminase
GLN synthetase

GDH
Glutaminase

Sources of Ammonia
1) Dietary protein
~60% dietary N are converted into urea in the liver
2) Enzymes of GIT bacteria
hydrolysis of GLN by intestinal bacteria. Bacteria also
degrade urea present in intestinal lumen.
3) Catabolism of endogenous amino acids
generating free NH3

emcrit.org/images/part1/ammonia%20produce.jpeg

putrescine
spermidine
spermine

Biomed Pharmacother 56:446-457, 2002

Glucagon
(why?)

Reactions of the urea cycle

Lippincotts Illustrated Reviews: Biochemistry, 2011

via shuttle

Urea cycle and TCA cycle interface

Citrin=aspartate-glutamate carrier
Citrin deficiency causes citrullinemia

http://www.ojrd.com/content/figures/1750-1172-7-32-1-l.jpg

Overall stoichiometry of the urea cycle

Aspartate + NH3 + CO2 + 3 ATP + H2O
urea + fumarate + 2ADP + AMP + 2Pi + PPi

Cytosolic &
mitochondrial
mitochondrial

cytosolic

Fum

Citrin deficiency causes

inefficient ASS activity and
limits urea cycle activity.
Mutation of ASS can also
occur, leading to type II
citrullinemia (and
hyperammonemia).
Saheki & Kobayahi. J Human Genet 47:333-341, 2002

Argininosuccinate lyase (ASL)

deficiency can lead to lifethreatening hyperammonemia.
Long-term complications may
include liver dysfunction,
cognitive deficit and
hypertension. Hypertension
may be due to secondary nitric
oxide deficiency.
Dietary therapy is key to
optimum health including
protein restriction and arginine
base supplementation. Liver
transplant may be required.
Metabolic fates of arginine. Arginine is derived from dietary sources, protein catabolism, and endogenous synthesis. The
citrullineNO cycle (green) is responsible for the regeneration of arginine in various tissues. Arginine serves as the
precursor for many biologically important molecules; a decrease in arginine may result in decreased production of
compounds for which it serves as a precursor. ADC, arginine decarboxylase; Arg1, arginase 1; ASA, argininosuccinic acid;
ASL, argininosuccinate lyase; ASS, argininosuccinate synthase; GABA, -amino butyric acid; GATM, glycine
amidinotransferase; NO, nitric oxide; NOS, NO synthase; OTC, ornithine transcarbamylase. Reproduced with permission
from publishers John Wiley and Sons.
Nagamani et al. Genetics in Medicine 14:501-507, 2012

Km of GLN Synthetase
for NH3 is low
(high affinity, low capacity)

escaping

Km of CPS I for NH3 is high

(low affinity, high capacity, not saturated easily.
Therefore, is the main channel of GLN metabolism)

Fail safe design to take care of body ammonia

emcrit.org/images/part1/ammonia%20produce.jpeg

Liver is the main organ of amino acid catabolism (except BCAA).

When liver function is impaired, the following nitrogen related metabolism may occur
which may play role in the development of hepatic encephalopathy or hepatic coma:
a)Reduced capacity of urea cycle, leading to elevated NH3
Ammonia is toxic to CNS because, more GLN will be formed leading to increase
osmotic pressure and cell swelling (edema). Also more GABA will be form from GLU (via
glutamic acid decarboxylase) which is an inhibitory neurotransmitter.
b) Elevated level of many amino acids including those which are neurotransmitter
precursors
AA precursor
PHE
TYR
TRP
HIS

Neurotransmitter
Norepinephrine (noradrenaline)
Epinephrine (adrenaline)
Dopamine
Serotonin
Histamine

Increase supply of precursors may disrupt normal neurotransmitter metabolism

and lead to formation of false neurotransmitters

lactulose*
Lactulose is a nondigestible disaccharide (fructose and galactose). Upon GIT
fermentation by bacteria produces SCFA therby lowering GIT pH and
transforms freely diffusible NH3 into NH4+ which cannot be diffuse back
into the blood.

Glutamine, Arginine and Nitric Oxide

Biomed Pharmacother 56: 427-438, 2002

Overloading of urea cycle could result from:

- NH4Cl consumption (accident)
- Increase protein degradation
- Arginine (ARG) deficient diet*
- High lysine (LYS) diet*
These conditions will lead to OROTIC ACIDURIA

which could be reversed by supplementing the diet with:

Citrulline, Ornithine or Arginine

* Often seen in chicken. Even more serious for cats because ARG is
essential.

Overloading of urea cycle

activates cytoplasmic
handling of GLN leading to
increase orotate.
?

Brosnan & Brosnan. J Nutr 137:1656S-1661S, 2007

http://www.chembio.uoguelph.ca/educmat/chm452/lectur30.htm

10

Biomed Pharmacother 56: 427-438, 2002

Pyrimidine synthesis in
cytosol from carbamoyl
asparate

(cytosine, thymine, uracil)

biochem4.okstate.edu/.../nucleotides.htm

11

Highlights for ARGININE synthesis

ARG is non-essential but could be conditionally essential
ARG synthesis involves the GIT and the kidney
Pyrroline-5-carboxylate synthetase is only found in GIT
Conversion of citrulline to ARG occurs in kidney
Requirement consideration cats & patients with massive
intestine resection.

Cats have very low intestinal activities of

Pyrroline-5-carboxylase synthetase &
Ornithine aminotransferase
As a consequence, citrulline supply is low. A single ARG-free
meal can result in hyperammonemia, convulsion and even death.

12

Role of the kidney

For ARG formation (~60% of net synthesis)
Takes up GLN for acid-base homeostasis
- sulfuric acids are formed from
dietary protein
- the hydrogen ions will be neutralized by
blood bicarbonate

Nutrition & Metabolism (2003, Blackwell Publishing)

OAT: ornithine animotransferase

Amino Acids 26:345-351, 2004

13

Biomed Pharmacother 56:439-445, 2002

http://www.djur.cob.lu.se/Cellprolif/Research/research_area_1.html

J Nutr Biochem 17: 571-588, 2006

Arterioscler Thromb Vasc Biol 28:s11-16, 2008

14

Amino acids as ingredients in supplements, functional

foods & beverages
Rationale:
- stimulate protein synthesis
- for various purposes including performance
enhancement (in sports), treatment of
bed sores, sarcopenia or after certain surgery.
Amino acids in single or combination:
BCAA
ARG
GLN
TAU
Observed safe level (OSL)
ARG 20 g/d
GLN 14 g/d

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