Metabolism of Amino Acids
Metabolism of Amino Acids
Metabolism of Amino Acids
15
Summary
1. The body proteins are in a dynamic state (degradation and synthesis) and there
is an active amino acid pool (100 g) maintained for this purpose.
2. The amino acids undergo transamination and deamination to liberate ammonia
for the synthesis of urea, the end product of protein metabolism.
3. Besides being present as structural components of proteins, amino acids
participate in the formation of several biologically important compounds.
4. Glycine is involved in the synthesis of creatine, heme, purines, glutathione
etc.
5. Phenylalanine is hydroxylated to tyrosine, which is a precursor for the
production of skin pigment (melanin), catecholamines (dopamine,
epinephrine and norepinephrine) and thyroid hormones (T3 and T4).
6. Tryptophan is converted to NAD+ and NADP+, the coenzymes of niacin,
serotonin (a neurotransmitter) and melatonin.
7. The active methionine (SAM) is a donor of methyl group (transmethylation)
for the synthesis of many biological compounds (epinephrine, choline,
methylcytosine etc.).
8. Many amino acids contribute to onecarbon fragments (formyl, formimino,
methylene etc.) for participation in onecarbon metabolism—which is mostly
under the control of tetrahydrofolate.
9. The carbon skeleton of amino acids is involved either in the synthesis of
glucose (glycogenic) or fat (ketogenic), or both-glucose and fat.
10. Many inborn errors (mostly due to enzyme defects) in amino acid
metabolism have been identified. These include phenylketonuria (defect-
phenylalanine hydroxylase), albinism (defect-tyrosinase), maple syrup urine
disease (defect−α-keto acid dehydrogenase) etc.
The amino acids speak :
“We transaminate and deaminate to liberate ammonia;
Proteins are the most abundant organic compounds and constitute a major
part of the body dry weight (10–12 kg in adults). They perform a wide variety of
static (structural) and dynamic (enzymes, hormones, clotting factors, receptors
etc.) functions. About half of the body protein (predominantly collagen) is
present in the supportive tissue (skeleton and connective) while the other half is
intracellular.
Proteins are nitrogen-containing macromolecules consisting of L-α-amino
acids as the repeating units. Of the 20 amino acids found in proteins, half can be
synthesized by the body (nonessential) while the rest have to be provided in the
diet (essential amino acids).
The proteins on degradation (proteolysis) release individual amino acids.
Amino acids are not just the structural components of proteins. Each one of the
20 naturally occurring amino acids undergoes its own metabolism and performs
specific functions. Some of the amino acids also serve as precursors for the
synthesis of many biologically important compounds (e.g. melanin, serotonin,
creatine etc.). Certain amino acids may directly act as neurotransmitters (e.g.
glycine aspartate, glutamate). Protein metabolism is more appropriately learnt
as metabolism of amino acids.
Amino acid pool
An adult has about 100 g of free amino acids which represent the amino acid
pool of the body. The amino acid pool may be an oversimplification of the facts,
since there is no single compartment—rather, several compartments exist.
Glutamate and glutamine together constitute about 50%, and essential amino
acids about 10% of the body pool (100 g). The concentration of intracellular
amino acids is always higher than the extracellular amino acids. Amino acids
enter the cells against a concentration gradient by active transport.
The amino acid pool of the body is maintained by the sources that contribute
(input) and the metabolic pathways that utilize (output) the amino acids
(Fig.15.1).
The details of general and specific metabolic reactions of amino acids are
described in the following pages.
Transamination
The transfer of an amino (−NH2) group from an amino acid to a keto acid is
known as transamination. This process involves the interconversion of a pair of
amino acids and a pair of keto acids, catalysed by a group of enzymes called
transaminases (recently, aminotransferases).
Mechanism of transamination
Transamination occurs in two stages (Fig.15.4)
FIG. 15.4 Mechanism of transamination—(A) Involvement of pyridoxal
phosphate (PLP) in the transfer of amino group, (B) Formation of enzyme-
PLP-Schiff base and amino acid-PLP-Schiff base.
Note that when the amino acid binds, enzyme separates.
Deamination
The removal of amino group from the amino acids as NH3 is deamination.
Transamination (discussed above) involves only the shuffling of amino groups
among the amino acids. On the other hand, deamination results in the liberation
of ammonia for urea synthesis. Simultaneously, the carbon skeleton of amino
acids is converted to keto acids. Deamination may be either oxidative or non-
oxidative.
Although transamination and deamination are separately discussed, they occur
simultaneously, often involving glutamate as the central molecule. For this
reason, some authors use the term transdeamination while describing the
reactions of transamination and deamination, particularly involving glutamate.
I Oxidative deamination
Oxidative deamination is the liberation of free ammonia from the amino group
of amino acids coupled with oxidation. This takes place mostly in liver and
kidney. The purpose of oxidative deamination is to provide NH3 for urea
synthesis and α-keto acids for a variety of reactions, including energy
generation.
The activity of L-amino acid oxidase is much low while that of D-amino acid
oxidase is high in tissues (mostly liver and kidney). L-Amino acid oxidase does
not act on glycine and dicarboxylic acids. This enzyme, due to its very low
activity, does not appear to play any significant role in the amino acid
metabolism.
II Non-oxidative deamination
Some of the amino acids can be deaminated to liberate NH3 without undergoing
oxidation
(a) Amino acid dehydrases : Serine, threonine and homoserine are the hydroxy
amino acids. They undergo non-oxidative deamination catalysed by
PLPdependent dehydrases (dehydratases).
(b) Amino acid desulfhydrases : The sulfur amino acids, namely cysteine and
homocysteine, undergo deamination coupled with desulfhydration to give
keto acids.
(c) Deamination of histidine : The enzyme histidase acts on histidine to liberate
NH3 by a non-oxidative deamination process.
Metabolism of ammonia
Ammonia is constantly being liberated in the metabolism of amino acids
(mostly) and other nitrogenous compounds. At the physiological pH, ammonia
exists as ammonium ( ) ion.
I Formation of ammonia
The production of NH3 occurs from the amino acids (transamination and
deamination), biogenic amines, amino group of purines and pyrimidines and by
the action of intestinal bacteria (urease) on urea.
Role of glutamine
Glutamine is a storehouse of NH3. It is present at the highest concentration (8
mg/dl in adults) in blood among the amino acids. Glutamine serves as a storage
and transport form of NH3. Its synthesis mostly occurs in liver, brain and muscle.
Ammonia is removed from the brain predominantly as glutamine. Glutamine is
freely diffusible in tissues, hence easily transported.
Glutamine synthetase (a mitochondrial enzyme) is responsible for the
synthesis of glutamine from glutamate and ammonia. This reaction is
unidirectional and requires ATP and Mg2+ ions.
Glutamine can be deaminated by hydrolysis to release ammonia by
glutaminase (Fig.15.8) an enzyme mostly found in kidney and intestinal cells.
V. Toxicity of ammonia
Even a marginal elevation in the blood ammonia concentration is harmful to the
brain. Ammonia, when it accumulates in the body, results in slurring of speech
and blurring of the vision and causes tremors. It may lead to coma and, finally,
death, if not corrected.
Hyperammonemia
Elevation in blood NH3 level may be genetic or acquired. Impairment in urea
synthesis due to a defect in any one of the five enzymes is described in urea
synthesis. All these disorders lead to hyperammonemia and cause hepatic coma
and mental retardation. The acquired hyperammonemia may be due to hepatitis,
alcoholism etc. where the urea synthesis becomes defective, hence NH3
accumulates.
Accumulation of NH3 shifts the equilibrium to the right with more glutamate
formation, hence more utilization of α-ketoglutarate. α-Ketoglutarate is a key
intermediate in TCA cycle and its depleted levels impair the TCA cycle. The net
result is that production of energy (ATP) by the brain is reduced. The toxic
effects of NH3 on brain are, therefore, due to impairment in ATP formation.
Urea cycle
Urea is the end product of protein metabolism (amino acid metabolism). The
nitrogen of amino acids, converted to ammonia (as described above), is toxic to
the body. It is converted to urea and detoxified. As such, urea accounts for 80–
90% of the nitrogen containing substances excreted in urine.
Urea is synthesized in liver and transported to kidneys for excretion in urine.
Urea cycle is the first metabolic cycle that was elucidated by Hans Krebs and
Kurt Henseleit (1932), hence it is known as Krebs-Henseleit cycle. The
individual reactions, however, were described in more detail later on by Ratner
and Cohen.
Urea has two amino (−NH2) groups, one derived from NH3 and the other
from aspartate. Carbon atom is supplied by CO2. Urea synthesis is a five-step
cyclic process, with five distinct enzymes. The first two enzymes are present in
mitochondria while the rest are localized in cytosol. The details of urea cycle are
described (Figs.15.9 and 15.10).
FIG. 15.9 Outline of urea cycle. (Note : In the synthesis of urea one
amino group comes from ammonium ion while the other is from aspartate;
carbon is derived from CO2. This is represented in colours.)
Disposal of urea
Urea produced in the liver freely diffuses and is transported in blood to kidneys,
and excreted. A small amount of urea enters the intestine where it is broken
down to CO2 and NH3 by the bacterial enzyme urease. This ammonia is either
lost in the feces or absorbed into the blood. In renal failure, the blood urea level
is elevated (uremia), resulting in diffusion of more urea into intestine and its
breakdown to NH3. Hyperammonemia (increased blood NH3) is commonly seen
in patients of kidney failure. For these patients, oral administration of antibiotics
(neomycin) to kill intestinal bacteria is advised.
FIG. 15.12 Interrelation between urea and tricarboxylic acid (TCA) cycle
(Depicted in blue colour).
Table 15.1
Metabolic defects in urea cycle
In some countries, estimations of BUN or NPN are used rather than blood
urea for assessing kidney function. The normal range for ratio of BUN to serum
creatinine is 10:1 to 15:1.
Table 15.2
A summary of the specialized products formed/contributed by
amino acids
Glycine
Glycine (Gly, G) is a nonessential, optically inactive and glycogenic (precursor
for glucose) amino acid. It is indispensable for chicks. The outline of glycine
metabolism is depicted in Fig.15.13. Glycine is actively involved in the
synthesis of many specialized products (heme, purines, creatine etc.) in the body,
besides its incorporation into proteins, synthesis of serine and glucose and
participation in onecarbon metabolism. Glycine is the most abundant amino acid
normally excreted into urine (0.5–1.0 g/g creatinine).
FIG. 15.13 Overview of glycine metabolism.
Glycine in proteins
Glycine is one among the commonest amino acids found in protein structure.
Being small and non-polar, glycine is mostly present in the interior structure of
protein. Collagen contains very high (about 30%) content of glycine.
Synthesis of glycine
Glycine is synthesized from serine by the enzyme serine hydroxymethyl
transferase which is dependent on tetrahydrofolate (THF). Glycine can also be
obtained from threonine, catalysed by threonine aldolase. Glycine synthase can
convert a onecarbon unit (N5, N10-methylene THF), CO2 and NH3 to glycine.
Degradation of glycine
Glycine undergoes oxidative deamination by glycine synthase to liberate ,
5 10
CO2 and onecarbon fragment as N , N -methylene THF. This provides a major
route for glycine breakdown in mammals. Glycine synthase is a multienzyme
complex and requires PLP, NAD+ and THF for its activity. This reaction is
reversible and, therefore, glycine can be generated from onecarbon unit
(methylene fragment of THF).
Glycine is reversibly converted to serine by THF dependent serine
hydroxymethyl transferase. Pyruvate produced from serine by serine
dehydratase, serves as a precursor for glucose. Serine is degraded to glyoxylate
which undergoes transamination to give back glycine. Glyoxylate is also
converted to oxalate, an excretory product and formate which enters onecarbon
pool (Fig.15.14).
Biomedical/clinical concepts
About 300–400 g of protein per day is constantly degraded and
synthesized in the human body.
The amino acids are mainly utilized for protein biosynthesis,
production of specialized products (creatine, porphyrin, amines,
purines, pyrimidines) and generation of energy.
Glutamate is the collection centre for the amino groups in the
biological system while glutamine is the storehouse of NH3. Free
NH3 can be liberated predominantly from glutamate.
Ammonia accumulation in blood is toxic to brain causing slurring
of speech, blurring of vision, tremors and even death. Mammals
convert NH3 to urea, a non-toxic excretory product. Metabolic
defects in urea cycle enzymes result in hyperammonemia.
Dietary consumption of a protein rich meal increases the level of
N-acetylglutamate in liver which enhances urea production.
Primary hyperoxaluria—a metabolic disorder due to a defect in
the enzyme glycine transaminase— is characterized by elevated
urinary oxalate and the formation of oxalate stones.
Blood urea estimation is commonly used to assess renal function.
Elevation of blood urea level (normal 10–40 mg/dl) is associated
with several disorders which may be prerenal (diabetic coma),
renal (acute glomerulonephritis) and post-renal (tumors or stones
in the urinary tract).
Estimation of serum creatinine (normal < 1 mg/dl) is considered
to be a more reliable indicator for the evaluation of kidney
function.
Synthesis of melanin
Melanin (Greek : melan—black) is the pigment of skin, hair and eye. The
synthesis of melanin occurs in melanosomes present in melanocytes, the
pigment-producing cells. Tyrosine is the precursor for melanin and only one
enzyme, namely tyrosinase (a copper-containing oxygenase), is involved in its
formation. Tyrosinase hydroxylates tyrosine to form 3,4-dihydroxyphenylalanine
(DOPA) (Fig.15.20). DOPA can act as a cofactor for tyrosinase. The next
reaction is also catalysed by tyrosinase in which DOPA is converted to
dopaquinone. It is believed that the subsequent couple of reactions occur
spontaneously, forming leucodopachrome followed by 5,6-dihydroxyindole. The
oxidation of 5, 6-dihydroxyindole to indole 5, 6-quinone is catalysed by
tyrosinase, and DOPA serves as a cofactor. This reaction, inhibited by tyrosine
regulates the synthesis of melanin. Melanochromes are formed from indole
quinone, which on polymerization are converted to black melanin.
Biosynthesis of catecholamines
The name catechol refers to the dihydroxylated phenyl ring. The amine
derivatives of catechol are called catecholamines.
Tyrosine is the precursor for the synthesis of catecholamines, namely
dopamine, norepinephrine (noradrenaline) and epinephrine (adrenaline).
The conversion of tyrosine to catecholamines occurs in adrenal medulla and
central nervous system involving the following reactions (Fig.15.22).
FIG. 15.22 Metabolism of tyrosine-synthesis of catecholamines
(dopamine, norepinephrine, epinephrine; PLP–pyridoxal phosphate).
Functions of catecholamines
Norepinephrine and epinephrine regulate carbohydrate and lipid metabolisms.
They stimulate the degradation of triacylglycerol and glycogen. They cause an
increase in the blood pressure. Dopamine and norepinephrine serve as
neurotransmitters in the brain and autonomous nervous system.
Biochemical basis
The exact biochemical cause of this disorder has not been identified. Parkinson's
disease is, however, linked with a decreased production of dopamine. The
disease is due to degeneration of certain parts of the brain (substantia nigra and
locus coeruleus), leading to the impairment in the synthesis of dopamine.
Treatment
Dopamine cannot enter the brain, hence its administration is of no use. DOPA
(levodopa or L-dopa) is used in the treatment of Parkinson's disease. In the brain,
DOPA is decarboxylated to dopamine which alleviates the symptoms of this
disorder. Unfortunately, dopamine synthesis occurs in various other tissues and
results in side-effects such as nausea, vomiting, hypretension etc. Administration
of dopa analogs—that inhibit dopa decarboxylase (in various tissues) but not
enter brain (due to blood-brain barrier)—are found to be effective. Carbidopa
and γ-methyl-dopa (dopa analogs) are administered along with dopa for the
treatment of Parkinson's disease.
Phenylketonuria
Phenylketonuria (PKU) is the most common metabolic disorder in amino acid
metabolism. The incidence of PKU is 1 in 10,000 births. It is due to the
deficiency of the hepatic enzyme, phenylalanine hydroxylase, caused by an
autosomal recessive gene. In recent years, a variant of PKU—due to a defect in
dihydrobiopterin reductase (relatively less)—has been reported. This enzyme
deficiency impairs the synthesis of tetrahydrobiopterin required for the action of
phenylalanine hydroxylase (See Fig.15.18). The net outcome in PKU is that
phenylalanine is not converted to tyrosine.
The name phenylketonuria is coined due to the fact that the metabolite
phenylpyruvate is a keto acid (C6H5CH2−CO−COO−) excreted in urine in high
amounts.
Diagnosis of PKU
PKU is mostly detected by screening the newborn babies for the increased
plasma levels of phenylalanine (PKU, 20–65 mg/ dl; normal 1–2mg/dl). This is
usually carried out by Guthrie test, which is a bacterial (Bacillus subtilis)
bioassay for phenylalanine. The test is usually performed after the baby is fed
with breast milk for a couple of days by testing elevated levels of phenylalanine.
Phenylpyruvate in urine can be detected by ferric chloride test (a green colour is
obtained). This test is not specific, since many other compounds give a false
positive test. Prenatal diagnosis of PKU can also be done by using cultured
amniotic cells.
Treatment of PKU
The maintenance of plasma phenylalanine concentration within the normal
range is a challenging task in the treatment of PKU. This is done by selecting
foods with low phenylalanine content and/or feeding synthetic amino acid
preparations, low in phenylalanine. Dietary intake of phenylalanine should be
adjusted by measuring plasma levels. Early diagnosis (in the first couple of
months of baby's life) and treatment for 4–5 years can prevent the damage to
brain. However, the restriction to protein diet should be continued for many
more years in life. Since the amino acid tyrosine cannot be synthesized in PKU
patients, it becomes essential and should be provided in the diet in sufficient
quantity.
In some seriously affected PKU patients, treatment includes administration of
5-hydroxytryptophan and dopa to restore the synthesis of serotonin and
catecholamines. PKU patients with tetrahydrobiopterin deficiency require
tetrahydrobiopterin supplementation.
Tyrosinemia type II
This disorder—also known as Richner-Hanhart syndrome, is due to a defect in
the enzyme tyrosine transaminase. The result is a blockade in the routine
degradative pathway of tyrosine. Accumulation and excretion of tyrosine and its
metabolites—namely p-hydroxyphenylpyruvate, p-hydroxyphenyllactate,
phydroxyphenylacetate, N-acetyltyrosine—and tyramine are observed.
Tyrosinemia type II is characterized by skin (dermatitis) and eye lesions and,
rarely, mental retardation. A disturbed self-coordination is seen in these patients.
Neonatal tyrosinemia
The absence of the enzyme p-hydroxyphenylpyruvate dioxygenase causes
neonatal tyrosinemia. This is mostly a temporary condition and usually responds
to ascorbic acid. It is explained that the substrate inhibition of the enzyme is
overcome by the presence of ascorbic acid.
Enzyme defect
The defective enzyme in alkaptonuria is homogentisate oxidase in tyrosine
metabolism (See Fig.15.19). Homogentisate accumulates in tissues and blood,
and is excreted into urine. Homogentisate, on standing, gets oxidized to the
corresponding quinones, which polymerize to give black or brown colour. For
this reason, the urine of alkaptonuric patients resembles coke in colour.
Biochemical manifestations
Homogentisate gets oxidized by polyphenol oxidase to benzoquinone acetate
which undergoes polymerization to produce a pigment called alkapton
(Fig.15.24). Alkapton deposition occurs in connective tissue, bones and various
organs (nose, ear etc.) resulting in a condition known as ochronosis. Many
alkaptonuric patients suffer from arthritis and this is believed to be due to the
deposition of pigment alkapton (in the joints), produced from homogentisate.
FIG. 15.24 Conversion of homogentisate to alkapton.
Diagnosis
Change in colour of the urine on standing to brown or dark has been the simple
traditional method to identify alkaptonuria.
The urine gives a positive test with ferric chloride and silver nitrate. This is
due to the strong reducing activity of homogentisate. Benedict's test—employed
for the detection of glucose and other reducing sugars—is also positive with
homogentisate.
Treatment
Alkaptonuria is not a dangerous disorder and, therefore, does not require any
specific treatment. However, consumption of protein diet with relatively low
phenylalanine content is recommended.
Albinism
Albinism (Greek: albino—white) is an inborn error, due to the lack of synthesis
of the pigment melanin. It is an autosomal recessive disorder with a frequency
of 1 in 20,000.
Biochemical basis
The colour of skin and hair is controlled by a large number of genes. About 150
genes have been identified in mice. The melanin synthesis can be influenced by
a variety of factors. Many possible causes (rather explanations) for albinism
have been identified
1. Deficiency or lack of the enzyme tyrosinase.
2. Decrease in melanosomes of melanocytes.
3. Impairment in melanin polymerization.
4. Lack of protein matrix in melanosomes.
5. Limitation of substrate (tyrosine) availability.
6. Presence of inhibitors of tyrosinase.
The most common cause of albinism is a defect in tyrosinase, the enzyme
most responsible for the synthesis of melanin (See Fig.15.20).
Clinical manifestations
The most important function of melanin is the protection of the body from sun
radiation. Lack of melanin in albinos makes them sensitive to sunlight. Increased
susceptibility to skin cancer (carcinoma) is observed. Photophobia (intolerance
to light) is associated with lack of pigment in the eyes. However, there is no
impairment in the eyesight of albinos.
Hypopigmentation
In some individuals, a reduced synthesis of melanin (instead of total lack) is
often observed. Hypopigmentation disorders may be either diffuse or localized.
A good example of diffuse hypopigmentation is oculocutaneous albinism
which is mostly due to mutations in the tyrosinase gene. The degree of
hypopigmentation depends on the type and severity of mutated genes.
Vitiligo and leukoderma are the important among the localized
hypopigmentation disorders. Vitiligo is an acquired progressive disease with loss
of pigmentation around mouth, nose, eyes and nipples. Leukoderma is
comparable with vitiligo, but lack of pigmentation usually begins with hands and
then spreads.
Greying of hair is due to lack of melanin synthesis which usually occurs as a
result of disappearance of melanocytes from the hair roots.
Tryptophan
Tryptophan (Trp, W) was the first to be identified as an essential amino acid. It
contains an indole ring and chemically it is α-amino β-indole propionic acid.
Tryptophan is both glucogenic and ketogenic in nature. It is a precursor for the
synthesis of important compounds, namely NAD+ and NADP+ (coenzymes of
niacin), serotonin and melatonin (Fig.15.25).
I Kynurenine pathway
This pathway mostly occurs in liver leading to oxidation of tryptophan and the
synthesis of NAD+ and NADP+ (Fig.15.26).
FIG. 15.26 Metabolism of tryptophan-kynurenine pathway (PLP–Pyridoxal
phosphate; QPRT–Quinolinate phosphoribosyl transferase; PRPP–
Phosphoribosyl pyrophosphate).
II Serotonin pathway
Serotonin or 5-hydroxytryptamine (5HT) is a neurotransmitter, synthesized from
tryptophan. Normally, about 1% of the tryptophan is converted to serotonin. The
production of 5HT occurs in the target tissues.
Synthesis of serotonin
In mammals, the largest amount of serotonin is synthesized in the intestinal
cells. The formation of serotonin is comparable with the production of
catecholamines. Tryptophan is first hydroxylated at 5th carbon by tryptophan
hydroxylase. This enzyme requires tetrahydrobiopterin as a cofactor. 5-
Hydroxytryptophan is decarboxylated by aromatic amino acid decarboxylase
(PLPdependent) to give serotonin (Fig.15.27).
Platelets contain high concentration of 5HT, the significance of which is not
clear. As such, platelets cannot synthesize serotonin.
Degradation of serotonin
Monoamine oxidase (MAO) degrades serotonin to 5-hydroxyindoleacetate
(5HIA) which is excreted in urine.
Functions of serotonin
Serotonin is a neurotransmitter and performs a variety of functions.
1. Serotonin is a powerful vasoconstrictor and results in smooth muscle
contraction in bronchioles and arterioles.
2. It is closely involved in the regulation of cerebral activity (excitation).
3. Serotonin controls the behavioural patterns, sleep, blood pressure and body
temperature.
4. Serotonin evokes the release of peptide hormones from gastrointestinal tract.
5. It is also necessary for the motility of GIT (peristalsis).
Diagnosis
The excretion of 5-hydroxy indole acetate in urine is tremendously elevated
(upto 500mg/day against normal <5 mg/day) in carcinoid syndrome. The
estimation of 5 HIA in urine is used for the diagnosis of this disorder. In general,
urine concentration of 5 HIA above 25 mg/day should be viewed with caution as
it may be suggestive of carcinoid syndrome. Sufficient precaution should,
however, be taken for sample collection. During the course of urine collection,
the patients should not ingest certain foods (banana, tomato etc.) that increase
urine 5 HIA.
Melatonin
Melatonin is a hormone, mostly synthesized by the pineal gland. Serotonin—
produced from tryptophan—is acted upon by serotonin N-acetylase (the rate
limiting enzyme), to give N-acetylserotonin. The latter undergoes methylation,
S-adenosylmethionine being the methyl group donor to produce melatonin or N-
acetyl 5-methoxyserotonin (Fig.15.27). The synthesis and secretion of melatonin
from pineal gland is controlled by light.
Functions of melatonin
1. Melatonin is involved in circadian rhythms or diurnal variations (24 hr
cyclic process) of the body. It plays a significant role in sleep and wake
process.
2. Melatonin inhibits the production of melanocyte stimulating hormone (MSH)
and adrenocorticotropic hormone (ACTH).
3. It has some inhibitory effect on ovarian functions.
4. Melatonin also performs a neurotransmitter function.
Hartnup's disease
This disorder was first described in the family of Hartnup, hence the name—
Hartnup's disease. It is a hereditary disorder of tryptophan metabolism. The
clinical symptoms include dermatitis, ataxia, mental retardation etc. Hartnup's
disease is characterized by low plasma levels of tryptophan and other neutral
amino acids and their elevated urinary excretion. Increased urinary output of
indoleacetic acid and indolepyruvic acid is also observed.
Pellagra-like symptoms are common in these patients. There is an impairment
in the synthesis of NAD+ and serotonin from tryptophan. Some authors (earlier)
attributed Hartnup's disease to a defect in the enzyme tryptophan pyrrolase. This,
however, does not appear to be true. Hartnup's disease is now believed to be due
to an impairment in the absorption and/or transport of tryptophan and other
neutral amino acids from the intestine, renal tubules and, probably brain. Some
more details on Hartnup's disease are given under digestion and absorption
(Chapter 8).
Metabolism of methionine
Methionine (or sulfur amino acids) metabolism may be divided into three parts.
1. Utilization of methionine for transmethylation reactions.
2. Conversion of methionine to cysteine and cystine.
3. Degradation of cysteine and its conversion to specialized products.
Transmethylation
The transfer of methyl group (−CH3) from active methionine to an acceptor is
known as transmethylation. Methionine has to be activated to S-
adenosylmethionine (SAM) or active methionine to donate the methyl group.
Synthesis of S-adenosylmethionine
The synthesis of S-adenosylmethionine occurs by the transfer of an adenosyl
group from ATP to sulfur atom of methionine (Fig.15.29). This reaction is
catalysed by methionine S-adenosyltransferase. The activation of methionine is
unique as the sulfur becomes a sulfonium atom (SAM is a sulfonium compound)
by the addition of a third carbon. This reaction is also unusual since all the three
phosphates of ATP are eliminated as pyrophosphates (PPi) and inorganic
phosphates (Pi). Three high energy phosphates (3 ATP) are consumed in the
formation of SAM.
Functions of S-adenosylmethionine
S-Adenosylmethionine is highly reactive due to the presence of a positive
charge. The enzymes involved in the transfer of methyl group are collectively
known as methyltransferases. S-Adenosylmethionine transfers the methyl group
to an acceptor and gets itself converted to S-adenosylhomocysteine. The loss of
free energy in this reaction makes the methyl transfer essentially irreversible. S-
Adenosylhomocysteine is hydrolysed to homocysteine and adenosine.
Homocysteine can be remethylated to methionine by N5-methyl tetrahydrofolate.
In this manner, methionine can be regenerated for reuse. It should be noted that
there is no net synthesis of methionine in the S-adenosyl-methionine cycle
(homocysteine, the precussor for methionine has to be derived from methionine).
Hence, methionine is an essential amino acid.
S-Adenosylmethionine (carbon fragment) is also involved in the synthesis of
polyamines (spermidine, spermine). The most important transmethylation
reactions are listed in Table 15.3.
Table 15.3
Transmethylation reactions
Significance of transmethylation
1. Transmethylation is of great biological significance since many compounds
become functionally active only after methylation.
2. Protein (amino acid residues) methylation helps to control protein turnover. In
general, methylation protects the proteins from immediate degradation.
3. In plants, S-adenosylmethionine is the precursor for the synthesis of a plant
hormone, ethylene, which regulates plant growth and development and is
involved in the ripening of fruits.
Synthesis of cysteine
Homocysteine formed from methionine is a precursor for the synthesis of
cysteine (Fig.15.30). Homocysteine condenses with serine to form cystathionine.
This reaction is catalysed by a PLPdependent cystathionine synthase. The
enzyme cystathioninase (PLPdependent) cleaves and deaminates cystathionine
to cysteine and α-ketobutyrate. The sum of the reactions catalysed by
cystathionine synthase and cystathioninase is a good example of
transsulfuration (transfer of sulfur from one compound to another). It should be
noted that only the sulfur atom of cysteine comes from homocysteine (originally
methionine) while the rest of the molecule is from serine.
FIG. 15.30 Synthesis of cysteine from methionine ( –For reactions of
methionine conversion to homocysteine see Fig. 15.29) ( –In cysteine
synthesis, only sulfur is obtained from homocysteine, the rest of the
molecule is from serine).
Degradation of cysteine
Cystine and cysteine are interconvertible by an NAD+-dependent cystine
reductase. Cysteine on decarboxylation produces mercaptoethanolamine which
is involved in the biosynthesis of coenzyme A from the vitamin pantothenic acid.
The enzyme cysteine dioxygenase oxidizes cysteine to cysteine sulfinate
which, on further oxidation, is converted to cysteic acid. The latter undergoes
decarboxylation to produce taurine which conjugates with bile acids. Cysteic
acid can also be degraded to pyruvate, which is glycogenic (Fig.15.31).
FIG. 15.31 Metabolism of cysteine and cystine.
Homocystinurias
Homocystinurias are a group of metabolic disorders characterized by the
accumulation and increased urinary excretion of homocysteine and S-
adenosylmethionine. Plasma concentration of methionine is increased.
Homocystinuria type I has been more thoroughly investigated. It is due to a
defect in the enzyme cystathionine synthase. Accumulation of homocysteine
results in various complications—thrombosis, osteoporosis and, very often,
mental retardation. Further, the deficiency of cystathionine is associated with
damage to endothelial cells which might lead to atherosclerosis. Two forms of
type I homocystinurias are known, one of them can be corrected with vitamin B6
supplementation (B6 responsive) while the other does not respond to B6. The
treatment includes consumption of diet low in methionine and high in cystine.
The patients of homocystinuria have high levels of homocysteine, and usually
die of myocardial infarction, stroke, or pulmonary embolism.
The other homocystinurias are associated with enzyme defects (as stated
below) in the conversion of homocysteine to methionine by remethylation.
Homocystinuria II
N5-N10—Methylene THF reductase.
Homocystinuria III
N5-N10—Methyl THFhomocysteine methyltransferase. This is mostly due to
impairment in the synthesis of methylcobalamin.
Homocystinuria IV
N5—Methyl THF homocysteine methyl transferase. This is primarily due to a
defect in the intestinal absorption of vitamin B12.
Onecarbon metabolism
Amino acid metabolism is particularly important for the transfer or exchange of
onecarbon units. The following onecarbon fragments are encountered in the
biological reactions, which constitute onecarbon pool
Methyl (−CH3)
Hydroxymethyl (−CH2OH)
Methylene (=CH2)
Methenyl (−CH=)
Formyl (−CH=O)
Formimino (−CH=NH)
[Note : It may be stated here that CO2 is also a onecarbon unit. Carbon
dioxide is involved (carboxylation) in many biochemical reactions, which are
dependent on biotin. For instance, conversion of pyruvate to oxaloacetate in
gluconeogenesis. Most of the authors, however, ignore CO2 as onecarbon unit
and do not even consider it worth mentioning. This would be unfair to CO2!]
Tetrahydrofolate (THF) is a versatile coenzyme that actively participates in
onecarbon metabolism. With regard to the transfer of methyl groups from S-
adenosylmethionine, vitamin B12 is also involved besides THF.
The onecarbon unit covalently binds with THF at position N5 or N10 or on both
N5 and N10 of pteroyl structure of folate. The details of different onecarbon units
binding with THF and the structures of THF derivatives are given under vitamin-
folic acid (Chapter 7).
The onecarbon metabolism is rather complex, involving many reactions. For
the sake of better understanding, it is divided into generation and utilization of
onecarbon units, and the role of methionine and vitamin B12.
Valine – glycogenic
Leucine – ketogenic
Isoleucine – glycogenic and ketogenic.
The first three metabolic reactions are common to the branched chain amino
acids (Fig.15.33).
FIG. 15.33 Summary of branched chain amino acid metabolism ( –
Defect in α-keto acid dehydrogenase results in maple syrup urine disease;
HMG CoA– β-Hydroxy β-methyl-glutaryl CoA; Valine is glycogenic, leucine
is ketogenic, while isoleucine is both).
Enzyme defect
Maple syrup urine disease is due to a defect in the enzyme branched chain α-
keto acid dehydrogenase. This causes a blockade in the conversion of α-keto
acids to the respective acyl CoA thioesters. The plasma and urine concentrations
of branched amino acids and their keto acids are highly elevated. This disease is
also known as branched chain ketonuria.
Histidine
The metabolism of histidine is important for the generation of onecarbon unit,
namely formimino group. The enzyme histidase acts on histidine to split off
ammonia. Urocanate formed in this reaction is acted upon by urocanase to
produce 4-imidazole 5-propionate. Imidazole ring of the product is cleaved by a
hydrolase to give N-formiminoglutamate (FIGLU). Tetrahydrofolate (THF)
takes up the formimino group to form N5-formimino THF, and glutamate is
liberated. Deficiency of folate blocks this reaction and causes elevated excretion
of FIGLU in urine. Histidine loading test is commonly employed to assess folate
deficiency.
Histidine, on decarboxylation, gives the corresponding amine—histamine.
Histamine regulates HCl secretion by gastric mucosa (Table 15.7). Excessive
production of histamine causes asthma and allergic reactions.
Table 15.4
Classification of amino acids based on the fate of carbon skeleton
Histidinemia
The frequency of histidinemia is 1 in 20,000. It is due to a defect in the enzyme
histidase. Histidinemia is characterized by elevated plasma histidine levels and
increased excretion of imidazole pyruvate and histidine in urine. Most of the
patients of histidinemia are mentally retarded and have defect in speech. No
treatment will improve the condition of the patients.
Proline
Proline is oxidized to pyrroline 5-carboxylate which undergoes a non-enzymatic
conversion to glutamate 5-semialdehyde. The latter is converted to glutamate
and then transaminated to α-ketoglutarate. The five carbons of proline are
converted to α-ketoglutarate.
Hyperprolinemia type I
It is due to a defect in the enzyme proline oxidase (proline dehydrogenase).
Another metabolic disorder—hyperprolinemia type II—associated with
hydroxyproline metabolism is also reported.
Arginine
Arginine is cleaved by arginase to liberate urea and produce ornithine. Ornithine
undergoes transamination of δ-amino group to form glutamate γ-semialdehyde
which is converted to glutamate. Hyperargininemia is an inborn error in
arginine metabolism due to a defect in the enzyme arginase.
Mechanism of action
Nitric oxide promotes the synthesis of cGMP. It is believed that some of the
actions of NO are mediated through cGMP and protein kinase G.
Agmatine
It is a derivative of arginine produced in the brain. Agmatine possesses
antihypertensive properties.
Lysine
Lysine is an essential amino acid. Cereal proteins are deficient in lysine. It does
not participate in transamination reactions. Some of the lysine residues in protein
structure are present as hydroxylysine, methyllysine or acetyllysine. ε-Amino
group of lysine, forming salt bridges is necessary for the maintenance of
structural conformation of protein.
Lysine is a ketogenic amino acid. The summary of lysine metabolism is
depicted in Fig.15.35.
FIG. 15.35 Summary of lysine metabolism.
Synthesis of carnitine
Some of the lysine residues in proteins are found in methylated form. The
methyl groups are obtained from active methionine (SAM). Such proteins on
degradation (by proteolysis) will release the methyllysines. The trimethyllysine
serves as a precursor for the synthesis of carnitine, a compound involved in the
transport of fatty acids to mitochondria for oxidation. It should be noted that free
lysine is not methylated, hence it will not be a substrate for carnitine formation.
Synthesis of carnitine from trimethyllysine is a 4-step reaction involving
oxidation, splitting off glycine residue, dehydrogenation and, finally, oxidation
(Fig.15.36).
Functions of GABA
It is one of the major inhibitory neurotransmitters in the brain. GABA regulates
the activity of neurons by discouraging the transmission signals. It is believed
that GABA opens chloride channels and increases the permeability of post-
synaptic membranes. Thus GABA functions as an inhibitory neurotransmitter.
Decreased GABA levels will cause convulsions.
Glutamine
Glutamine is a versatile amino acid. Ammonia is temporarily stored in the form
of glutamine. Glutamine is freely diffusible and, hence, easily transported. The
synthesis and degradation of glutamine are described (See Fig.15.8).
Glutamine is the donor of nitrogen atoms for purine and pyrimidine synthesis.
It is the chief source of ammonia in kidneys. The NH3 production is elevated in
acidosis to maintain acid-base balance. Glutamine also takes part in
conjugation reactions.
The diagnostic importance of the enzyme AST has already been described
(Chapter 6). Aspartate has certain important functions (Fig.15.39).
FIG. 15.39 Overview of aspartate and asparagine metabolism.
1. It donates one amino group for the synthesis of urea (the other amino group in
urea directly comes from ammonia).
2. Aspartate forms a connecting link between urea cycle and TCA cycle (via
oxaloacetate).
3. It is utilized for the synthesis of purines (N1 and NH2 at 6th position) and
pyrimidines (N3, C4 C5 and C6 atoms).
4. Malate-aspartate shuttle is important for the transfer of reducing equivalents
(NADH) from the cytosol to mitochondria (Refer Fig.11.13).
Asparagine is synthesized from aspartate by a synthetase in an irreversible
ATP-dependent reaction. Asparaginase hydrolyses asparagine and liberates
ammonia (Fig.15.40). These reactions are comparable to glutamine synthesis
and its breakdown.
FIG. 15.40 Synthesis of asparagine and its conversion to aspartate (Note
: The reactions are independent and irreversible).
Alanine
The nonessential amino acid alanine performs two important functions in the
body— incorporation into proteins and participation in transamination and NH3
transport.
As already discussed, ammonia is toxic to the body, hence it cannot be
transported in free form. Glutamate and glutamine shoulder the major burden of
ammonia transport. Alanine is also important in this regard. In the peripheral
tissues (most predominantly—muscle), pyruvate produced in glycolysis gets
converted to alanine (by transamination) and is transported to liver. Pyruvate can
be regenerated from alanine in liver and the pyruvate so produced serves as a
precursor for glucose. Amino group is diverted for transamination or urea
formation. This is an alanine-pyruvate shuttle for carrying nitrogen to be
reutilized or converted to urea.
It is recently (2008) reported that people with higher levels of alanine in
urine have increased risk for higher blood pressure.
The amino acid β-alanine is a constituent of the vitamin pantothenic acid, and
thus the coenzyme A.
Serine
Serine is a nonessential glycogenic amino acid. As described in glycine
metabolism, serine and glycine are interconvertible. Serine can be synthesized
from the intermediates of glycolysis (3-phosphoglycerate). The metabolic
reactions of serine are described hereunder (Fig.15.41)
FIG. 15.41 Overview of serine metabolism.
Threonine
Threonine is an essential hydroxy amino acid. It is glycogenic and does not
participate in transamination reactions. Threonine is often a carrier of phosphate
group in the protein structure. The outline of threonine metabolism is depicted in
Fig.15.42.
Table 15.6
Inborn errors of amino acid metabolism
Biomedical/clinical concepts
Melanin—the pigment of skin, hair and eyes—is produced from
tyrosine. Lack of melanin synthesis (mostly due to a deficiency of
tyrosinase) causes albinism.
Parkinson's disease—a common disorder of the elderly—is linked
with decreased synthesis of dopamine. It is characterized by
muscular rigidity, tremors, lethargy etc.
Phenylketonuria, due to a defect in the enzyme phenylalanine
hydroxylase, is characterized by failure of growth, seizures and
mental retardation (low IQ).
Alkaptonuria causes the accumulation of homogentisate which
undergoes oxidation followed by polymerization to produce the
pigment alkapton. Deposition of alkapton in connective tissue,
causes ochronosis which is associated with arthritis.
Serotonin, an excitatory neurotransmitter, is synthesized from
tryptophan. Psychic stimulant drugs (iproniazid) elevate serotonin
levels while depressant drugs (LSD) decrease.
Malignant carcinoid syndrome, a tumor of argentaffin cells of
gastrointestinal tract, is characterized by tremendously increased
production of serotonin. This disorder can be diagnosed by the
elevated levels of 5-hydroxyindoleacetate in urine.
Melatonin, produced from serotonin, is involved in circadian
rhythms or diurnal variations, i.e., maintenance of body's
biological clock.
Homocysteine has been implicated as a risk factor in the onset of
coronary heart diseases.
Histidine loading test, characterized by elevated excretion of N-
formiminoglutamate (FIGLU) is commonly employed to assess
the deficiency of the vitamin, folic acid.
Nitric oxide (NO), synthesized from arginine, is involved in
several biological functions— vasodilation, platelet aggregation,
neurotransmission and bactericidal action. NO is used in the
treatment of chronic obstructive pulmonary disease (COPD).
γ-Aminobutyric acid (GABA), produced from glutamate, is an
inhibitory neurotransmitter. Low levels of GABA result in
convulsions.
The carbon skeleton of amino acids may be converted to glucose
(glycogenic) or fat (ketogenic), besides being responsible for the
synthesis of nonessential amino acids.
Polyamines (spermine, putrescine) are involved in the synthesis of
DNA, RNA and proteins and, thus, they are essential for cell
growth and differentiation.
Amino aciduria
The term amino aciduria is generally used to indicate the urinary excretion of
amino acids. It is frequently associated with defects in amino acid metabolism.
Most of the amino acidurias manifest in mental retardation.
Table 15.7
Amino acids and their derivatives as neurotransmitters
Biogenic amines
In general, the decarboxylation of amino acids or their derivatives results in the
formation of amines.
A summary of the biogenic amines derived from different amino acids and
their major functions are given in Table 15.8.
Table 15.8
A summary of the biogenic amines and their functions
Polyamines
Polyamines (Greek: poly—many) possess multiple amino groups. Putrescine,
spermine and spermidine are the biologically important polyamines. Spermine
and spermidine were originally detected in human semen (sperms), hence they
are so named.
Biosynthesis
Ornithine and S-adenosylmethionine are the precursors for polyamine synthesis.
It should, however, be noted that only the four-carbon moiety of SAM (not the
methyl group) is involved in polyamine formation. Ornithine decarboxylase acts
on ornithine to split off CO2 and produce putrescine (Fig.15.44). The enzyme
ornithine decarboxylase has the shortest half-life (about 10 minutes) among the
known mammalian enzymes. It regulates polyamine synthesis. The activity of
this enzyme is increased by hormones like corticosteroids, testosterone and
growth hormone.
Degradation of polyamines
The enzyme polyamine oxidase (of liver peroxisomes) oxidizes spermine to
spermidine and then to putrescine. Spermidine and putrescine are excreted in
urine in a conjugated form, as acetylated derivatives. Some amount of putrescine
is also oxidized to NH3 and CO2.