1.

Introduction
2. Aminocidopathies
3. Amino Acid Analysis
i. Building blocks of proteins
ii. Growth, repair and maintenance of cells
} Nutritionally essential amino acids is supplied
by diet (e.g. phenylalanine, lysine, etc.)
} Phenylalanine is a precursor for tyrosine,
the monoamine signaling
molecules dopamine, norepinephrine (noradr
enaline), and epinephrine (adrenaline), and
the skin pigment melanin.
} Phenylalanine is found naturally in the breast
milk of mammals.
} Lysine is important for proper growth, and it
plays an essential role in the production of
carnitine, a nutrient responsible for converting
fatty acids into energy and helping to lower
cholesterol. Lysine appears to help the body
absorb calcium, and it plays an important role in
the formation of collagen, a substance important
for bones and connective tissues including skin,
tendon, and cartilage.
} Not produced by our body
} Some amino are produced in the body
(e.g. tyrosine, glutamine, etc.)
} Tyrosine (abbreviated as Tyr or Y) or 4-
hydroxyphenylalanine, is one of the 22 amino
acids that are used
by cells to synthesize proteins.
} Glutamine (abbreviated as Gln or Q) is one of the
20 amino acids encoded by the standard genetic
code. It is not recognized as an essential amino
acid, but may become conditionally essential in
certain situations, including intensive athletic
training or certain gastrointestinal disorders
q Class of inherited errors of metabolism
q Enzymes defects that inhibits the body’s
ability to metabolize certain amino acids
i. PKU vi. Homocystinuria
ii. Tyrosinemia vii. Citrullinemia
iii. Alkaptonuria viii. Arginosuccinic
Aciduria
iv. MSUD
ix. Cystinuria
v. Isovaleric Acidemia
Metabolism of Phenylalanine and
Tyrosine
PAH
} Absence of phenylalanine hydroxylase (PAH)
} Musty odor of urine
} is an autosomal recessive metabolic genetic
disorder characterized by a mutation in the gene for
the hepatic enzymephenylalanine hydroxylase (PAH),
rendering it nonfunctional.
} This enzyme is necessary to metabolize the amino
acid phenylalanine (Phe) to the amino acid tyrosine.
When PAH activity is reduced, phenylalanine
accumulates and is converted
into phenylpyruvate (also known as phenylketone),
which can be detected in the urine.
} Mental retardation and microencephaly
} - deficient enzyme: phenylalanine
hydroxylase

} Low tetrahydrobiopterin – cofactor

} Increased amino acid
} =phenylalanine
} = phenyllactic acid
} = phenylpyruvate (phenylketone)
} Disease :
} Mental retardation, microcephaly, musty odor
urine

} Lab test :
} Guthrie test and microfluorometric assay
i. Phenylketonuria
§ Laboratory Tests
1. Guthrie test
§ Semi quantitative bacterial inhibition assay
§ Uses phenylalanine to facilitate bacterial
growth (B. subtilis and β-2-
thienylalanine).
Metabolism of Phenylalanine and
Tyrosine
} ↓ Tyrosine aminotransferase

Deficient enzyme: Tyrosine aminotransferase

Increased amino acid: Tyrosine
Disease manifestation:
Mental retardation, phtophobia, eye pain
} Type II tyrosinemia is caused by a deficiency of the
enzyme tyrosine aminotransferase .
} Tyrosine aminotransferase is the first in a series of
five enzymes that converts tyrosine to smaller
molecules, which are excreted by the kidneys or used
in reactions that produce energy.
} This form of the disorder can affect the eyes, skin,
and mental development.
} Symptoms often begin in early childhood and include
excessive tearing, abnormal sensitivity to light
(photophobia), eye pain and redness, and painful skin
lesions on the palms and soles. About half of
individuals with type II tyrosinemia are also mentally
challenged.
} Deficient enzyme:
} ↓ 4-hydroxyphenylpyruvate dioxygenase

} Increased AA: p-hydroxyphenylpyruvic acid (PHPAA)

} dISEASE :
} Mild metntal retardation,
} rare disorder caused by a deficiency of the
enzyme 4-hydroxyphenylpyruvate dioxygenase.
} This enzyme is abundant in the liver, and smaller
amounts are found in the kidneys. It is one of a
series of enzymes needed to break down
tyrosine. Specifically, 4-hydroxyphenylpyruvate
dioxygenase converts a tyrosine byproduct
called 4-hydroxyphenylpyruvate to homogentisic
acid.
} Characteristic features of type III tyrosinemia
include mild mental retardation, seizures, and
periodic loss of balance and coordination
(intermittent ataxia). Type III tyrosinemia is very
rare; only a few cases have been reported.
} Lack of homogentisate oxidase
} ↑ homogentesic acid in urine
} Alkaptonuria (black urine disease or alcaptonuria) is a
rare inherited genetic disorder
of phenylalanine and tyrosine metabolism.
} This is an autosomal recessive condition that is due
to a defect in the enzyme homogentisate 1,2-
dioxygenase , which participates in the degradation
of tyrosine.
} As a result, homogentisic acid and its oxide,
called alkapton, accumulate in the blood and are
excreted in urine in large amounts (hence -uria).
Excessive homogentisic acid causes damage
to cartilage (ochronosis, leading to osteoarthritis)
and heart valves as well as precipitating as kidney
stones.
} Disease :
} Ochronosis (bluish black discoloration of tissues)
} Dark spot on sclera (white part of eye)
} Deposition of pigment in cartilage of nose, eyes,
tendons (hips,joints) – arthritis-like degeneration

} Urine turns brownish-black upon exposure w air

LAB TEST: FERRIC CHLORIDE TEST
- Test presence for phenol
- Urine+ FeCl = urine turns BLACK (+)
} ↓ fumarylacetoacetate hydrolase
} HIGH Fumarylacetoacetate

} Disease:
} Cabbage (cirrhosis or liver cancer)
} kIdney failure
} Cabbage-like odor
} Type 1 Tyrosinemia, also known as hepatorenal
tyrosinemia, is the most severe form of tyrosinemia. It is
caused by a deficiency of the enzymefumarylacetoacetate
hydrolase and p-hydroxyphenylpyruvic acid oxidase
} The primary effects are progressive liver and kidney
dysfunction. The liver disease causes cirrhosis, conjugated
hyperbilirubinemia, elevated AFP, hypoglycemia and
coagulation abnormalities. This can lead
to jaundice, ascites and hemorrhage.
} There is also an increased risk of hepatocellular
carcinoma.
} The kidney dysfunction presents as Fanconi syndrome:
Renal tubular acidosis, hypophosphatemia and
aminoaciduria. Cardiomyopathy, neurologic and
dermatologic manifestations are also possible.
 Phenylalanine hydroxylase

Phenylketonur
ia
Tyrosine Tyrosinemi
aminotransferase a II
Tyrosinemia III

4-hydroxyphenylpyruvate
dioxygenase

Alkaptonur Homogentisate
ia oxidase

Fumarylacetoacetate Tyrosinemia
hydrolase I
} metabolic disorder caused by a deficiency of
the branched-chain alpha-keto acid
dehydrogenase complex (BCKDC),

} leading to a buildup of the branched-chain

amino acids (leucine, isoleucine, and valine) and
their toxic by-products (ketoacids) in the blood
and urine.

} Symptoms:
} Mental retardation ,
} burnt sugar odor of urine, skin, and breath
vi. Laboratory Tests
1. Modified Guthrie test
§ uses branched chain α-ketoacid to facilitate
bacterial growth (containing B. subtilis and
4-azaleucine- inhibitor).
2) Microfluorometric assay
} Deficiency of isovaleryl-CoA dehydrogenase
} ↑ Isovaleric acid
} ↑ Leucine
} (for leucine
} metabolism)

} Sweaty feet odor

} Can damage the brain
} and CNS
} Some asymptomatic
} Ms/Ms chromatography
} Lack of cystathionine β- synthetase (cbs gene
} ↑ Homocysteine Methionine & Met &
Homocysteine
} Symptom
} Near sightedness
} Dissociation of
lens in eye
-Osteoporosis
} also known as cystathionine beta synthase
deficiency or CBS deficiency
} is an inherited disorder of the metabolism of
the amino acid methionine,
} It is an inherited autosomal recessive trait
} which means a child needs to inherit a copy
of the defective gene from both parents to be
affected.
} his defect leads to a multisystemic disorder
of the connective tissue, muscles, CNS,
and cardiovascular system.
§ Laboratory Tests
1. Modified Guthrie test
-bacterial inhibition test
- Uses methionine to facilitate bacterial
growth (containing B. subtilis and L-
methionine sulfoximime)

Confirmatory: MS/MS & HPLC

} 1) Citrullinemia
} 2) Arginosuccinic aciduria
} Citrullinemia is an autosomal recessive urea
cycle disorder that causes ammonia and other
toxic substances to accumulate in
the blood. Since the substances also
accumulate in the urine,
ix. Citrullinemia – urea cycle disorder= excess
urea = toxic in CNS brain damage
i. Type I citrullinemia
§ Lack of arginonosuccinic acid synthetase (ASS)
§ Increase – citrulline and ammonia
§ Disease – CNS , Brain damage, death (untreated)

Urea
Cycle
Mutation of ASS gene = accumulate citrulline & ammonia
} usually becomes evident in the 1st few days of life. Affected
infants typically appear normal at birth,
} but as ammonia builds up in the body, they develop a
} lack of energy (lethargy),
} poor feeding, (lack of apetite)
} vomiting, seizure, coma (toxic ammonia)
} loss of consciousness.
} severe brain damage, deah (not treated_

} These medical problems can be life-threatening in many

cases. A milder form of type I citrullinemia is less common in
childhood or adulthood. Some people with gene mutations
that cause type I citrullinemia never experience signs and
symptoms of the disorder.
Citrullinemia
A) Type II citrullinemia
-Mutation of the gene that encodes
for protein citrin (SLC25A13)

Citrin Urea
- Transport molecule inside cell used
Cycle
in production & breakdown of
- simple sugars, protein production,
urea cycle
} The symptoms of type II citrullinemia
} usually appear during adulthood and mainly
affect the central nervous system.
} Characteristic features include confusion,
abnormal behaviors (such as aggression,
irritability, and hyperactivity), seizures, and
coma.
} These symptoms can be life-threatening, and
are known to be triggered by certain
medications, infections, and alcohol intake in
people with this type.
 § Lack of argininosuccinic acid lyase (ASL)
§ Increase ammonia, citrulline, arginonosuccinate
x. Resp for Ammonia to urea conversion = nitrogen excess
xi. Lethargy, unwillingness to eat
xii. Disease
xiii. - CNS symptoms
xiv. (seizure and coma)

Urea
Cycle
} is an inherited disorder that causes the
accumulation of argininosuccinic acid (also
known as "ASA") in the blood and urine.
} Some patients may also have an elevation of
ammonia, a toxic chemical, which can affect
the nervous system.
} Argininosuccinic aciduria may become
evident in the first few days of life because of
high blood ammonia, or later in life
presenting with "sparse" or "brittle" hair,
developmental delay, and tremors.
} An infant with argininosuccinic aciduria may seem
lethargic or be unwilling to eat, have poorly-
controlled breathing rate or body temperature,
} experience seizures or unusual body movements, or
go into a coma.
} Complications from argininosuccinic aciduria may
include developmental delay and mental retardation.
} Progressive liver damage, skin lesions, and brittle
hair may also be seen. Immediate treatment and
lifelong management (following a strict diet and
using appropriate supplements) may prevent many of
these complications.
§ Defect in amino acid transport system
Mutation in SLS3AL and SLC7A9 gene – for protein synthesis
§ Inadequate reabsorption of cystine in the kidneys kaya
mataas cysteine
§ Disease : stones in kidney, ureter, bladder
§ Lab test :
§ Cyanide nitroprusside test (+) red purple color
§ False (+): Homocysteine sa sample
} Symptoms cystinuria
} - since cysteine if saturated pwede maalis sa
urine:
} Stones
} Hematuria
} Flank pain
} Uti

Treatment : increase fluid intake for stones

U Phenylalanine hyroxylase Phenylalanine
Fumarylacetoacetate
osinemia I Fumarylacetoacetate
hydrolase
osinemia II Tyrosine aminotranferase Tyrosine
p-
4-Hydroxyphenylpyruvate
osinemia III Hydroxyphenylpyruvic
oxidase
acid
Homogentisic acid
aptonuria Homogentisate oxidase
(HGA)
Branched-chain Leucine, Isoleucine,
UD
α-ketoacid decarboxylase Valine
valeric Isovaleryl-CoA
Leucine, Isovaleric acid
demia dehydrogenase
Homocysteine,
mocystinuria Cystathionine-β synthetase
Methionine
Arginosuccinic acid
q Diagnosis
i. 6-8 hours fasting
ii. Collected in heparin tube with the plasma
iii. Deproteinization performed within 30
minutes
iv. Screening by TLC stained with ninhydrin
(blue)
v. Separated and quantitated by Ion
exchange chromatography and HPLC
reversed-phase system or capillary
electrophoresis
} - blood sample: drawin after at least 6-8 hrs
fasting
} Collect in heparin tube
} Promptly separate plasma from cells
(centrifuge agad)
} Deproteinization done within 30 min
} Samples run immediately
} If test cannot be done, store at -20 to -40 C
} (freeze)
} Urinary amino acid analysis – can be perform on
random specimen for screening purposes
} Quantitation -24hr urine specimen preserved with
thymol or organic solvent
} Amniotic fluid analyze

} Amino acid screening – thin layer chromatography