METABOLISM OF NUCLEIC ACID

Nucleotides Metabolism
Ribonucleoside and deoxyribonucleoside phosphates (nucleotides) are essential for all cells.
Without them, neither DNA nor RNA can be produced and, therefore, proteins cannot be
synthesized or cells proliferate.
Nucleotides also serve as carriers of activated intermediates in the synthesis of some
carbohydrates, lipids, and proteins, and are structural components of several essential
coenzymes, for example, coenzyme A, FAD, NAD+, and NADP+. Nucleotides, such as cyclic AMP
(cAMP) and cyclic GMP (cGMP), serve as second messengers in signal transduction pathways.
Finally, nucleotides are also important regulatory compounds for many of the pathways of
intermediary metabolism, inhibiting or activating key enzymes.
The purine and pyrimidine bases found in nucleotides can be synthesized de novo, or can be
obtained through salvage pathways that allow the reuse of the preformed bases obtained from
normal cell turnover or from the diet.
Purine and pyrimidine nucleotides are synthesized in vivo at rates consistent with physiologic
need. Intracellular mechanisms, sense and regulate the pool sizes of nucleotide triphosphates
(NTPs), which rise during growth or tissue regeneration when cells are rapidly dividing.

BIOSYNTHESIS OF PURINE NUCLEOTIDES

Three processes contributes to purine nucleotide biosynthesis. These in order of decreasing
importance are:
(1) synthesis from amphibolic intermediates (synthesis de novo)
(2) phosphoribosylation of purines
(3) phosphorylation of purine nucleosides.

De novo synthesis of Purine nucleotide

The atoms of the purine ring are contributed by a number of compounds, including amino acids
(aspartic acid, glycine, and glutamine), CO2, and N10-formyltetrahydrofolate. The purine ring
is constructed by a series of reactions that add the donated carbons and nitrogens to a
preformed ribose 5-phosphate produced by the HMP pathway.
The pathway for the de novo synthesis of Purine nucleotide could be broken down into stages
and steps as follows:-
1- Synthesis of 5-phosphoribosyl-1 -pyrophosphate (PRPP)
Synthesis of PRPP from ATP and ribose 5-phosphate is catalyzed by PRPP synthetase (ribose
phosphate pyrophosphokinase. This enzyme is activated by inorganic phosphate (Pi) and
inhibited by purine nucleotides (end-product inhibition).

2- Synthesis of 5'-phosphoribosylamine
Synthesis of 5'-phosphoribosylamine from PRPP and glutamine. The amide group of glutamine
replaces the pyrophosphate group attached to carbon 1 of PRPP. The enzyme,
glutamine:phosphoribosyl pyrophosphate amidotransterase, is inhibited by the purine 5'-
nucleotides AMP, GMP, and IMP (the end products of the pathway). This is the committed step
in purine nucleotide biosynthesis. The rate of the reaction is also controlled by the intracellular
concentration of the substrates glutamine and PRPP.
3- Synthesis of inosine monophosphate, the "parent" purine nucleotide
The next nine steps in purine nucleotide biosynthesis leading to the synthesis of IMP (whose
base is hypoxanthine). This pathway requires four ATP molecules as an energy source. Two
steps in the pathway require N10-formyltetrahydrofolate.
4-Conversion of IMP to AMP and GMP
The conversion of IMP to either AMP or GMP uses a two-step, energy-requiring pathway.
The synthesis of AMP requires GTP as an energy source, whereas the synthesis of GMP requires
ATP. Also, the first reaction in each pathway is inhibited by the end product of that pathway.
This provides a mechanism for diverting IMP to the synthesis of the species of purine present in
lesser amounts. If both AMP and GMP are present in adequate amounts, the de novo pathway
of purine synthesis is turned off at the amidotransferase step.

5. Conversion of nucleoside monophosphates to nucleoside diphosphates and triphosphates

Nucleoside diphosphates (NDP) are synthesized from the corresponding nucleoside

monophosphates (NMP) by base-specific nucleoside monophosphate kinases. These kinases
do not discriminate between ribose or deoxyribose in the substrate. Generally, ATP is the
source of the transferred phosphate, because it is present in higher concentrations than the
other nucleoside triphosphates. Adenylate kinase is particularly active in liver and muscle,
where the turnover of energy from ATP is high. Its function is to maintain an equilibrium among
AMP, ADP, and ATP. Nucleoside diphosphates and triphosphates are interconverted by
nucleoside diphosphate kinase—an enzyme that, unlike the monophosphate kinases, has broad
specificity.
SALVAGE PATHWAY FOR PURINES BIOSYNTHESIS

Purines that result from the normal turnover of cellular nucleic acids, or that are obtained from
the diet and not degraded, can be reconverted into nucleoside triphosphates and used by the
cells. This is referred to as the "salvage pathway" for purines synthesis.
Conversion of purines, their ribonucleosides, and their deoxyribonucleosides to
mononucleotides require far less energy than de novo synthesis. The more important
mechanism involves phosphoribosylation by PRPP of a free purine to form a Purine 5’-
mononucleotide.

1. Conversion of Purine Bases to Nucleotides:

Two enzymes are involved: adenine phosphoribosyltransferase (APRT) and hypoxanthine-
guanine phosphoribosyltransferase (HPRT). Both enzymes use PRPP as the source of the ribose
5-phosphate group. The release of pyrophosphate makes these reactions irreversible.
2. Conversion of Purine nucleoside to Nucleotides:

A second salvage mechanism involves phosphoryl transfer from ATP to a purine ribonucleoside
(PuR). Adenosine kinase catalyzes phosphorylation of adenosine and deoxyadenosine to AMP
and dAMP, and deoxycytidine kinase phosphorylates deoxycytidine and 2’-deoxyguanosine to
dCMP and dGMP.

Liver, the major site of purine nucleotide biosynthesis, provides purines and purine nucleosides
for salvage and utilization by tissues incapable of their biosynthesis. For example, human brain
has a low level of PRPP amidotransferase and hence depends in part on exogenous purines.
Erythrocytes and polymorphonuclear leukocytes cannot synthesize 5-phosphoribosylamine

SYNTHESIS OF DEOXYRIBONUCLEOTIDES

The nucleotides required for DNA synthesis are 2'-deoxyribonucleotides, which are produced
from ribonucleoside diphosphates by the enzyme ribonucleotide reductase.

Ribonucleotide reductase
Ribonucleotide reductase (ribonucleoside diphosphate reductase) is a multisubunit enzyme (two
identical B1 subunits and two identical B2 subunits) that is specific for the reduction of
nucleoside diphosphates (ADP, GDP, CDP, and UDP) to their deoxy-forms (dADP, dGDP, dCDP,
and dUDP). The immediate donors of the hydrogen atoms needed for the reduction of the 2'-
hydroxyl group are two sulfhydryl groups on the enzyme itself, which, during the reaction, form
a disulfide bond.
Regeneration of reduced enzyme:
In order for ribonucleotide reductase to continue to produce deoxyribonucleotides, the
disulfide bond created during the production of the 2'-deoxy carbon must be reduced. The
source of the reducing equivalents is thioredoxin—a peptide coenzyme of ribonucleotide
reductase. Thioredoxin contains two cysteine residues separated by two amino acids in the
peptide chain. The two sulfhydryl groups of thioredoxin donate their hydrogen atoms to
ribonucleotide reductase, in the process forming a disulfide bond.
Regeneration of reduced thioredoxin: Thioredoxin must be converted back to its reduced form
in order to continue to perform its function. The necessary reducing equivalents are provided
by NADPH + H+, and the reaction is catalyzed by the enzyme thioredoxin reductase.
DEGRADATION OF PURINE NUCLEOTIDES

Degradation of dietary nucleic acids occurs in the small intestine, where a family of pancreatic
enzymes hydrolyzes the nucleotides to nucleosides and free bases. Inside cells, purine
nucleotides are sequentially degraded by specific enzymes, with uric acid as the end product of
this pathway.
Mammals other than primates oxidize uric acid further to allantoin, which, in some animals
other than mammals, uric acid is further degraded to urea or ammonia

A. Degradation of dietary nucleic acids in the small intestine

Ribonucleases and deoxyribonucleases, secreted by the pancreas, hydrolyze RNA and DNA
primarily to oligonucleotides. Oligonucleotides are further hydrolyzed by pancreatic
phosphodiesterases, producing a mixture of 3'- and 5'-mononucleotides. A family of
nucleotidases removes the phosphate groups hydrolytically, releasing nucleosides that may be
absorbed by the intestinal mucosal cells, or be further degraded to free bases before uptake.
Dietary purines and pyrimidines are not used to a large extent for the synthesis of tissue nucleic
acids. Instead, the dietary purines are generally converted to uric acid by intestinal mucosal
cells. Most of the uric acid enters the blood, and is eventually excreted in the urine. For this
reason, individuals with a tendency toward gout should be careful about consuming foods such
as organ meats, red meat, sardines, or dried beans, which contain high amounts of nucleic
acids.

Formation of uric acid

A summary of the steps in the production of uric acid and genetic diseases associated with
deficiencies of specific degradative enzymes are as follows

[1] An amino group is removed from AMP to produce IMP, or from adenosine to produce
inosine (hypoxanthine-ribose) by AMP or adenosine deaminase.
[2] IMP and GMP are converted into their nucleoside forms inosine and guanosine—by the
action of 5'-nucleotidase.

[3] Purine nucleoside phosphorylase converts inosine and guanosine into their respective
purine bases, hypoxanthine and guanine.

[4] Guanine is deaminated to form xanthine.

[5] Hypoxanthine is oxidized by xanthine oxidase to xanthine, which is further oxidized by

xanthine oxidase to uric acid, the final product of human purine degradation which is excreted
in the urine.

DISORDERS OF PURINE BIOSYNTHESIS

1. Lesch-Nyhan syndrome: This syndrome is an X-linked, recessive disorder associated

with virtually complete deficiency of HPRT. This deficiency results in inability to salvage
hypoxanthine or guanine, from which excessive amounts of uric acid are produced. In addition,
the lack of this salvage pathway causes increased PRPP levels and decreased IMP and GMP
levels. As a result, glutamine:phosphoribosylpyrophosphate amidotransferase (the committed
step in purine synthesis) has excess substrate and decreased inhibitors available, and de novo
purine synthesis is increased. The combination of decreased purine reutilization and increased
purine synthesis results in the production of large amounts of uric acid, making the Lesch-
Nyhan syndrome a severe, heritable form of gout. Patients with Lesch-Nyhan syndrome tend to
produce urate kidney stones. In addition, characteristic neurologic features of the disorder
include self-mutilation and involuntary movements.

Tophaceous gout, in which tophi (nodular masses of monosodium urate crystals) are deposited
in the soft tissues of the body. The deposition of needle-shaped monosodium urate crystals
initiates an inflammatory process involving the infiltration of granulocytes that phagocytize the
urate crystals. This process generates oxygen metabolites that damage tissue, resulting in the
release of lysosomal enzymes that evoke an inflammatory response. In addition, lactate
production in the synovial tissues increases, resulting in a decrease in pH that fosters further
deposition of urate crystals. A definitive diagnosis requires aspiration and examination of
synovial fluid using polarized light microscopy to confirm the presence of monosodium urate
crystals.

a. Primary gout: In most patients, gout is caused by the underexcretion of uric acid due to
defective renal secretion. However, overproduction of uric acid may occur because of an
inherited abnormality in the enzymes of purine metabolism. This is defined as "primary gout."
For example, several X-linked mutations have been identified in the PRPP synthetase gene
that result in the enzyme having either an increased Vmax for the production of PRPP, a lower
Km for ribose 5-phosphate, or a decreased sensitivity to its Purine nucleotide inhibitors. In each
case, purine production is increased, resulting in elevated levels of plasma uric acid.
Lesch-Nyhan syndrome also causes hyperuricemia, as a result of the decreased salvage of
hypoxanthine and guanine bases.

b. Secondary gout: This form of gout is caused by a variety of disorders and lifestyles, for
example, in patients with chronic renal insufficiency, those undergoing chemotherapy, those
with myeloproliferative disorders, and those who consume excessive amounts of alcohol or
purine-rich foods. Gout can also be an adverse effect of seemingly unrelated metabolic
diseases, such as von Gierke disease or fructose intolerance.

Von Gierke’s Disease

Purine overproduction and hyperuricemia in von Gierke’s disease (glucose-6-phosphatase
deficiency) occurs secondary to enhanced generation of the PRPP precursor ribose 5-
phosphate. An associated lactic acidosis elevates the renal threshold for urate, elevating total
body urates.

2. Adenosine deaminase deficiency:

Adenosine deaminase (ADA) is expressed in the cytosol of all cells however, humans
lymphocytes have the highest activity of this enzyme. A deficiency of ADA results in an
accumulation of adenosine, which is converted to its ribonucleotide or deoxyribonucleotide
forms by cellular kinases. As dATP levels rise, ribonucleotide reductase is inhibited, thus
preventing the production of all deoxyribose-containing nucleotides. Consequently, cells cannot
make DNA and divide. In its most severe form, this autosomal recessive disorder causes severe
combined immunodeficiency disease (SCID), involving a lack of both T cells and B cells.

Treatment for gout:

Acute attacks are treated with colchicines to decrease movement of granulocytes into the
affected area, and with anti-inflammatory drugs, such as aspirin, to provide
pain relief. Most therapeutic strategies for gout involve lowering the uric acid level below the
saturation point, thus preventing the deposition of urate crystals.
Uricosuric agents, such as probenecid or sulfinpyrazone, 5 are used in most patients with gout
as a result of underexcretion of uric acid.
Allopurinol—an inhibitor of uric acid synthesis—is more toxic, and is reserved for patients with
hyperuricemia as a result of overproduction of uric acid. Allopurinol is converted in the body to
oxypurinol, which inhibits xanthine oxidase, resulting in an accumulation of hypoxanthine and
xanthine compounds more soluble than uric acid and, therefore, less likely to initiate an
inflammatory response.

PYRIMIDINE SYNTHESIS AND DEGRADATION

Unlike the synthesis of the purine ring, in which the ring is constructed on a preexisting ribose
5-phosphate, the pyrimidine ring is synthesized before being attached to ribose 5-phosphate,
which is donated by PRPP. The sources of the atoms in the pyrimidine ring are glutamine, C02,
and aspartic acid. Glutamine and aspartic acid are therefore required for both purine and
pyrimidine synthesis

A. Synthesis of carbamoyl phosphate

The regulated step of this pathway in mammalian cells is the synthesis of carbamoyl phosphate
from glutamine and C02, catalyzed by carbamoyl phosphate synthetase II (CPS II). CPS II is
inhibited by UTP (the end-product of the pathway, which can be converted into the other
pyrimidine nucleotides), and is activated by ATP and PRPP.

B. Synthesis of orotic acid

The second step in pyrimidine synthesis is the formation of carbamoylaspartate, catalyzed by
aspartate transcarbamoylase. The pyrimidine ring is then closed hydrolytically by
dihydroorotase. The resulting dihydroorotate is oxidized to produce orotic acid (orotate)
The enzyme, dihydroorotate dehydrogenase that produces orotate, is located inside the
mitochondria. All other reactions in pyrimidine biosynthesis are cytosolic.
The first three enzymes in this pathway (CPS II, aspartate transcarbamoylase, and
dihydroorotase) are all domains of the same polypeptide chain. This is an example of a
multifunctional or multicatalytic polypeptide that facilitates the ordered synthesis of an
important compound.
C. Formation of a pyrimidine nucleotide
The completed pyrimidine ring is converted to the nucleotide orotidine 5'-monophosphate
(OMP) in the second stage of pyrimidine nucleotide synthesis . PRPP is the ribose 5-phosphate
donor. The enzyme orotate phosphoribosyltransferase produces OMP and releases
pyrophosphate, thereby making the reaction biologically irreversible.

Orotidine 5'-monophosphate (OMP), the parent pyrimidine mononucleotide, is converted to

uridine monophosphate (UMP) by orotidylate decarboxylase, which removes the acidic
carboxyl group. Orotate phosphoribosyltransferase and orotidylate decarboxylase are also
domains of a singlepolypeptide chain called UMP synthase.
Orotic aciduri:
A rare genetic defect—is caused by a deficiency of the bifunctional enzyme (UMP synthase)
Orotate phosphoribosyltransferase and orotidylate decarboxylase, resulting in orotic acid in the
urine.

D. Synthesis of uridine triphosphate and cytidine triphosphate

Cytidine triphosphate (CTP) is produced by amination of UTP by CTP synthetase. The nitrogen is
provided by glutamine—another example of a reaction in nucleotide biosynthesis in which this
amino acid is required.
E. Syntheisis of thymidine monophosphate from dUMP

dUMP is converted to dTMP by thymidylate synthase, which uses N5,N10-methylene

tetrahydrofolate as the source of the methyl group. This is an unusual reaction in that
tetrahydrofolate (THF) contributes not only a carbon unit but also two hydrogen atoms from
the pteridine ring, resulting in the oxidation of THF to dihydrofolate (DHF).
Salvage of pyrimidines biosynthesis
Few pyrimidine bases are salvaged in human cells. However, the pyrimidine nucleosides uridine
and cytidine can be salvaged by uridine-cytidine kinase, deoxycytidine can be salvaged by
deoxycytidine kinase, and thymidine can be salvaged by the enzyme thymidine kinase. Each of
these enzymes catalyzes the phosphorylation of a nucleoside(s) utilizing ATP, and forming UMP,
CMP, dCMP, and TMP.
While mammalian cells reutilize few free pyrimidines, “salvage reactions” convert the
ribonucleosides uridine and cytidine and the deoxyribonucleosides thymidine and
deoxycytidine to their respective nucleotides. ATP dependent phosphoryltransferases (kinases)
catalyze the phosphorylation of the nucleoside diphosphates 2’-deoxycytidine, 2’-
deoxyguanosine, and 2’-deoxyadenosine to their corresponding nucleoside triphosphates. In
addition, orotate phosphoribosyltransferase (reaction 5, Figure 34–7), an enzyme of pyrimidine
nucleotide synthesis, salvages orotic acid by converting it to orotidine monophosphate (OMP).

Degradation of pyrimidine nucleotides

Unlike the purine rings, which are not cleaved in human cells, the pyrimidine ring can be
opened and degraded to highly soluble structures, such as p-alanine, and p-aminoisobutyrate,
which can serve as precursors of acetyl CoA and succinyl CoA, respectively.

OVERPRODUCTION OF PYRIMIDINE CATABOLITES IS ONLY RARELY ASSOCIATED WITH

CLINICALLY SIGNIFICANT ABNORMALITIES
Since the end products of pyrimidine catabolism are highly water-soluble, pyrimidine
overproduction results in few clinical signs or symptoms. In hyperuricemia associated with
severe overproduction of PRPP, there is overproduction of pyrimidine nucleotides and
increased
excretion of -alanine. Since N5,N10-methylene- tetrahydrofolate is required for thymidylate
synthesis, disorders of folate and vitamin B12 metabolism result in deficiencies of TMP.
Orotic Acidurias
The orotic aciduria that accompanies Reye’s syndrome probably is a consequence of the
inability of severely damaged mitochondria to utilize carbamoyl phosphate, which then
becomes available for cytosolic overproduction of orotic acid. Type I orotic aciduria reflects a
deficiency
of both orotate phosphoribosyltransferase and orotidylate decarboxylase ; the rarer type II
orotic aciduria is due to a deficiency only of orotidylate decarboxylase.

Deficiency of a Urea Cycle Enzyme Results in Excretion of Pyrimidine Precursors

Increased excretion of orotic acid, uracil, and uridine accompanies a deficiency in liver
mitochondrial ornithine transcarbamoylase. Excess carbamoyl phosphate exits to the cytosol,
where it stimulates pyrimidine nucleotide biosynthesis. The resulting mild orotic aciduria is
increased by high nitrogen foods.

Drugs May Precipitate Orotic Aciduria

Allopurinol an alternative substrate for orotate phosphoribosyltransferase, competes with
orotic acid. The resulting nucleotide product also inhibits orotidylate decarboxylase, resulting
in orotic aciduria and orotidinuria. 6-Azauridine, following conversion to 6-azauridylate, also
competitively inhibits orotidylate decarboxylase, enhancing excretion of orotic acid and
orotidine.
[Note: Mycophenolic acid (MPA) is a potent, reversible, uncompetitive inhibitor of inosine
monophosphate dehydrogenase that is being used successfully in preventing graft rejection.
It blocks the de novo formation of guanosine monophosphate (GMP, see Figure 22.8), thus
depriving rapidly proliferating cells, including T and B cells, of a key component of nucleic
acids.]
REGULATION OF NUCLEOTIDE BIOSYNTHESIS

Purine & Pyrimidine Nucleotide Biosynthesis Are Coordinately Regulated

Purine and pyrimidine biosynthesis parallel one another mole for mole, suggesting coordinated
control of their biosynthesis. Several sites of cross-regulation characterize purine and
pyrimidine nucleotide biosynthesis. The PRPP synthase reaction (reaction 1, Figure 34–2), which
forms a precursor essential for both processes, is feedback-inhibited by both purine and
pyrimidine nucleotides

Inhibitors of thymidylate synthase include thymine analogs such as 5-fluorouracil, which serve
as successful antitumor agents.6 5-Fiuorouracil is metabolically converted to 5-FdUMP, which
becomes permanently bound to the inactivated thymidylate synthase; for this reason, the drug
is called a "suicide" inhibitor. DHF can be reduced to THF by dihydrofolate reductase (see Figure
28.3, p. 372), an enzyme that is inhibited in the presence of drugs such as methotrexate. By
decreasing the supply of THF, these folate analogs not only inhibit purine synthesis (see Figure
22.7), but, by preventing methylation of dUMP to dTMP, they also lower the cellular
concentration of this essential component of DNA. DNA synthesis is, therefore, inhibited and
cell growth slowed. Because of their ability to slow the replication of DNA by decreasing the
availability of nucleotide precursors, drugs such as those described above are used to decrease
the growth rate of cancer cells.

The intermediates and reactions for conversion of -D-ribose 5’-phosphate to Inosine

monophosphate (IMP). Separate branches then lead to AMP and GMP. Subsequent phosphoryl
transfer from ATP converts AMP and GMP to ADP and GDP. Conversion of GDP to GTP involves
a second phosphoryl transfer from ATP, whereas conversion of ADP to ATP is achieved primarily
by oxidative phosphorylation.

AMP & GMP Feedback-Regulate PRPP Glutamyl Amidotransferase

Since biosynthesis of IMP consumes glycine, glutamine, tetrahydrofolate derivatives, aspartate,
and ATP, it is advantageous to regulate purine biosynthesis. The major determinant of the rate
of de novo purine nucleotide biosynthesis is the concentration of PRPP, whose pool size
depends on its rates of synthesis, utilization, and degradation. The rate of PRPP synthesis
depends on the availability of ribose 5-phosphate and on the activity of PRPP synthase, an
enzyme sensitive to feedback inhibition by AMP, ADP, GMP, and GDP.

Two mechanisms regulate conversion of IMP to GMP and AMP. AMP and GMP feedback-inhibit
adenylosuccinate synthase and IMP dehydrogenase, respectively. Furthermore, conversion of
IMP to adenylosuccinate en route to AMP requires GTP, and conversion of xanthinylate (XMP)
to GMP requires ATP. This cross-regulation between the pathways of IMP metabolism thus
serves to decrease synthesis of one purine nucleotide when there is a deficiency of the other
nucleotide. AMP and GMP also inhibit hypoxanthine-guanine phosphoribosyltransferase, which
converts hypoxanthine and guanine to IMP and GMP and GMP feedback- inhibits PRPP
glutamyl amidotransferase.

Methotrexate Blocks Reduction of Dihydrofolate

The only reaction of pyrimidine nucleotide biosynthesis that requires a tetrahydrofolate
derivative. The methylene group of N5,N10-methylene- tetrahydrofolate is reduced to the
methyl group that is transferred, and tetrahydrofolate is oxidized to dihydrofolate. For further
pyrimidine synthesis to occur, dihydrofolate must be reduced back to tetrahydrofolate, a
reaction catalyzed by dihydrofolate reductase. Dividing cells, which must generate TMP and
dihydrofolate, thus are especially sensitive to inhibitors of dihydrofolate reductase such as the
anticancer drug methotrexate.

Gene Expression & Enzyme Activity Both Are Regulated

The activities of the first and second enzymes of pyrimidine nucleotide biosynthesis are
controlled by allosteric regulation. Carbamoyl phosphate synthase II is inhibited by UTP and
purine nucleotides but activated by PRPP. Aspartate transcarbamoylase is inhibited by CTP but
activated by ATP. In addition, the first three and the last two enzymes of the pathway are
regulated by coordinate repression and derepression.
.

SUMMARY
• Ingested nucleic acids are degraded to purines and pyrimidines. New purines and pyrimidines
are formed from amphibolic intermediates and thus are dietarily nonessential.
• Several reactions of IMP biosynthesis require folate derivatives and glutamine. Consequently,
antifolate drugs and glutamine analogs inhibit purine biosynthesis.
• Oxidation and amination of IMP forms AMP and GMP, and subsequent phosphoryl transfer
from ATP forms ADP and GDP. Further phosphoryl transfer from ATP to GDP forms GTP. ADP is
converted to ATP by oxidative phosphorylation. Reduction of NDPs forms dNDPs.
• Hepatic purine nucleotide biosynthesis is stringently regulated by the pool size of PRPP and by
feedback inhibition of PRPP-glutamyl amidotransferase by AMP and GMP.
• Coordinated regulation of purine and pyrimidine nucleotide biosynthesis ensures their
presence in proportions appropriate for nucleic acid biosynthesis and other metabolic needs.
• Humans catabolize purines to uric acid (pKa 5.8), present as the relatively insoluble acid at
acidic pH or as its more soluble sodium urate salt at a pH near neutrality. Urate crystals are
diagnostic of gout. Other disorders of purine catabolism include Lesch- Nyhan syndrome, von
Gierke’s disease, and hypouricemias.
• Since pyrimidine catabolites are water-soluble, their overproduction does not result in clinical
abnormalities. Excretion of pyrimidine precursors can, however, result from a deficiency of
ornithine transcarbamoylase because excess carbamoyl ph phosphate is available for pyrimidine
biosynthesis.