Explain the fate of carbon skeleton and nitrogen group of

amino acids.

Explain the ways of transport of nitrogen from various parts of
the body to the liver

Describe the urea cycle and the enzymes involved in
production of urea in the liver

Define and classify Hyperammonemias . List the enzymes
deficient in various hyperammonemias and its clinical
features

Amino group NH
3
Formation of urea
Carbon skeletons Formation of Glucose and
Ketone bodies.
FATE OF THE CARBON SKELETONS
Carbon skeletons are used for
energy
Glucogenic: TCA cycle
intermediates or Pyruvate
(Gluconeogenesis)
Ketogenic: Acetyl CoA,
Acetoacetyl CoA, or
Acetoacetate
PROTEINS
Gastric juice ( acidity denatures proteins )
Intestinal enzymes hydrolyze

AMINO ACIDS
Amino acid transporters Na+Amino acid symporter (can take
up di and tri peptides )
UPTAKE
DEFECTS :-
1)Hartnups disease Long neutral amino acid transporter
defect( Trp is not taken up Pellagra like symptoms as Trp
Niacin is not formed )
2)Cystinuria Basic amino acid transporter defect that also
transports cysteineUrinary stones .Cystine forms Cystine by
disulfide linkage (less soluble Stones )
OVERVIEW OF AMINO ACID METABOLISM
ENVIRONMENT ORGANISM


Ingested
protein



Bio-
synthesis




Protein




AMINO
ACIDS




Nitrogen
NH
3





Carbon
skeletons



Urea



Degradation
(required)
1
2 3
a
b

Purines
Pyrimidines
Porphyrins

c c

Used for
energy


pyruvate
-ketoglutarate
succinyl-CoA
fumarate
oxaloacetate


acetoacetate
acetyl CoA

(glucogenic) (ketogenic)
Recycling
Affects central nervous system

1. Alkalization of intracellular compartment

2. Disrupts oxidative phosphorylation ATP depletion

3. Increased glutamate in Brain

4. Decreased Neurotransmitters GABA convulsions

5. Cerebral edema
Symptoms of AMMONIA toxicity
Flapping Tremor (Asterixis)
( Correlate flapping tremor later on with Liver
failure in Clinical medicine )
Slurred Speech
Blurred Vision
COMA Death
Ammonotelic Fishes
Lots of water available

Uricotelic Reptiles and birds
Birds have to keep minimum body weight for flight

Ureotelic Mammals

Body Proteins
Amino acids
Catabolism
UREA
25%
80-85%
Sources of Amino Acids :
Exogenous Diet

Endogenous
1. Breakdown of muscle
protein
2. Biosynthesis from
intermediates of citric acid
cycle.

Utilization of Amino acids:
Synthesis of New proteins

Formation of Nucleotides

Formations of Porphyrins
and Catecholamines

Production of energy and
Ammonia.
1) Reutilization: Glutamate and Glutamine are involved in
recycling of amino acids.
Glutamate + Ammonia Glutamine
Glutamine
Synthase
ATP ADP
They are secreted by the peripheral tissues in form of
glutamine which is taken up by hepatocytes where the NH3 is
re-used for amino acid and nucleotide synthesis
Glutamine Glutamate + Ammonia
Glutaminase
Two important reactions are involved in fixing
ammonia back to amino acids:

1. Reductive Amination:
2. Amino Transferases:

All non-essential amino acids except for
tyrosine and cysteine are derived and are
dependent on transamination from glutamate.
1. Reductive Amination :
Alpha keto glutarate +
Ammonia
Glutamate
NAD(P)H NAD(P)
2. Transamination reaction:
Amino acid-1 Alpha keto acid-1
Amino acid-2 Alpha keto acid-2
Transaminase
Glutamate
dehydrogenase
PLP
Well balanced polarity
(Quite uncharged
because of amide
nitrogen yet sufficiently
soluble in plasma No
transporter required )


Non-Toxic

AMMONIA
ASPARTATE
UREA BIOSYNTHESIS IS DIVIDED INTO 4 STAGES:-

1. TRANSAMINATION

2. OXIDATIVE DEAMINATION

3. AMMONIA TRANSPORT

4. REACTIONS OF THE UREA CYCLE

DEF :- THE TRANSFER OF THE ALPHA-AMINO GROUP
FROM ONE AMINO ACID TO A KETO ACID , RESULTING IN
FORMATION OF A NEW AMINO ACID AND
CORRESPONDING KETO ACID .

E.G :- REACTION CATALYZED BY ALANINE AMINOTRANSFERASE
ALANINE PYRUVATE
(AMINO ACID ) (CORRESPONDING KETO ACID )


-KETOGLUTARATE GLUTAMATE
(KETO ACID ) (NEW AMINO ACID)

ALL TRANSAMINASES
REQUIRE PLP (VIT B6)
KA
AA
Corresponding KA
New AA
TRANSAMINATION
Aminoacids:

1. Transamination
2. Deamination
3. Transulfuration
4. Glutaminase
5. Gastrointestinal tract bacteria

Degradation of Amino sugars
Monoamine Oxidase
Pyrimidine catabolism
Nitrogen part is toxic. Excreted in the form of
either :
1. Ammonia charged and alkaline. Excreted as
ammonium ion in urine (3%)

2. Urea Neutral molecule Non toxic ( 80-85%)

3. Creatinine (3-4%) constant in urine ( 1% of Creatine
every day)

4. Uric acid from Purines only !

Inter organ exchange of amino acids

The Glucose alanine cycle


Kaplan lecture notes USMLE step 1

Kaplan lecture notes USMLE step 1
Very
important
NH
3
removal
mechanism
( esp BRAIN)

Kaplan lecture notes USMLE step 1
WHY ALANINE ? NOT GLUTAMATE DIRECTLY

2
ND
NITROGEN
1
ST
NITROGEN
(AST)
(ALT)
Aminoacids:

1. Transamination
2. Deamination
3. Transulfuration
4. Glutaminase
5. Gastrointestinal tract bacteria

Degradation of Amino sugars
Monoamine Oxidase
Pyrimidine catabolism
Nitrogen is Excreted in the form of either :
1. Ammonia charged and alkaline. Excreted as
ammonium ion in urine (3%)

2. Urea Neutral molecule Non toxic ( 80-85%)

3. Creatinine (3-4%) constant in urine ( 1% of
Creatine every day)

4. Uric acid from Purines only !

Inter organ exchange of amino acids in
post absorptive state (FASTING)

Inter organ exchange of amino acids in
absorptive state (after feeding)

The Glucose alanine cycle

ALT
(Transamination)
ALT
(Transamination)
Well balanced polarity
(Quite non polar
because of amide
nitrogen yet sufficiently
soluble in plasma No
transporter required )


Non-Toxic

AMMONIA
ASPARTATE
Ammonia + Bicarbonate + ATP
Carbomyl Phosphate
Citrulline
Arginosuccinate
Arginine
CPS -1
Ornithine Transcarbomylase
Arginosuccinate synthase
Arginosuccinate Lyase
Arginase
Ornithine
Aspartate
NAG N-acetyl Glutamate
High protein Diet
Cytoplasm
Mitochondria
Fumarate
Urea
activator
TCA cycle
Oxaloacetate

Acetyl Co-A + Glutamate N-acetyl Glutamate
N-acetyl glutamate synthase
(NAGS)
(NAG)
N-acetyl glutamate is the allosteric activator of Carbomyl
phosphate synthase-1.
Arginine
Urea cycle disorders
Hyperammonemia
Encephalopathy
Respiratory alkalosis
VOMITING
AVOIDANCE OF HIGH PROTEIN FOODS
INTERMITTENT ATAXIA
LETHARGY
SEVERE MENTAL RETARDATION
Symptoms of AMMONIA toxicity
Tremor
( Correlate flapping tremor later on with Liver
failure in Clinical medicine )
Slurred Speech
Blurred Vision
COMA Death
Hyperammonemia type -1
Autosomal recessive

Defect in CPS- 1

1 in 200,000

No orotic aciduria

Cerebral Oedema ,
coma and death .

Hyperammonemia type -2
X-linked recessive

Defect in OTC

MC urea cycle defect

Orotic aciduria present

Cerebral oedema , coma and
death.

CAUSE OF OROTIC ACIDURIA Increased
Carbamoyl phosphate spills out from
mitochondia to cytosolPyrimidine synthesis
Orotic acid
3. Citrullinemia :
Defect in arginosuccinate synthase
Citrullinuria
Autosomal recessive

4. Arginosuccinic aciduria:
Defect in arginosuccinate lyase
Arginosuccinic acid blood, CSF, Urine

5. Hyperargininemia :
Diet without arginine
Defect in arginase enzyme
:
Limit protein intake

Decrease bacterial source of ammonia Antibiotics
(Like Neomycin, Azithromycin)and Lactulose
(purgative )
Replace intermediates of urea cycle Arginine
Citrulline, Aspartate
Remove excess ammonia Hemodialysis, sodium
benzoate, phenyl acetate

Very Important
Very Important
Lactulose Acidification conversion to NH4+ and induction of
Purgation
Mainstay

Gut sterilization :-Neomycin/Azithromycin other antibiotics
Very Important

Combination of Sodium benzoate and
Phenylacetate/Phenylbutyrate

Sodium benzoate + Glycine Hippuric acid excreted

Phenylacetate phenylacetyl glutamine excreted. (Phenylacetate conjugates
with glutamine to form phenylacetylglutamine, which is excreted by the kidneys)
Rarely used
Source http://biocadmin.otago.ac.nz
MCQ 1
Select the CORRECT answer .
The first reaction in the degradation of the majority of
common amino acids involves participation of :
A. NAD +
B. Pyridoxal Phosphate
C. Thiamine Pyrophosphate(TPP)
D. FAD
E. NAD and TPP
MCQ 2
After thorough investigations a man is diagnosed
with orotic aciduria . To find out the cause of
orotic aciduria which of the following
investigations will you prefer?
A. ALP levels
B. vitamin b12 assay
C. FIGLU excretion assay
D. Peripheral smear
E. serum bilirubin