28

Chapter
THE METABOLISM OF PURINES
AND PYRIMIDINES

The purine and pyrimidine bases (Fig. 28.1) are constitu- PURINE SYNTHESIS STARTS WITH
ents of nucleotides and nucleic acids. The ribonucleotides RIBOSE-5-PHOSPHATE
adenosine triphosphate (ATP), guanosine triphosphate
De novo synthesis of purines is most active in the liver,
(GTP), uridine triphosphate (UTP), and cytidine triphos-
which exports the bases and nucleosides to other tis-
phate (CTP) are present in millimolar concentrations
sues. Most tissues have a limited capacity for de novo
in the cell. They serve important coenzyme functions in
purine synthesis, although they can synthesize the
addition to being precursors of RNA synthesis. The deox-
nucleotides from externally supplied bases.
yribonucleotides deoxyadenosine triphosphate (dATP),
The pathway of purine biosynthesis is shown in
deoxyguanosine triphosphate (dGTP), deoxycytidine
Figure 28.2. It starts with ribose-5-phosphate, a product
triphosphate (dCTP), and deoxythymidine triphosphate
of the pentose phosphate pathway (see Chapter 22).
(dTTP) are present in micromolar concentrations and
In the reactions of the pathway, all of them cytoplasmic,
are required only for DNA replication and DNA repair.
the purine ring system is built up step by step, with C-1
Their cellular concentrations are highest during S phase
of ribose-5-phosphate used as a primer.
of the cell cycle.
The first enzyme, 5-phosphoribosyl-1-pyrophosphate
Dietary nucleic acids and nucleotides are digested to
(PRPP) synthetase, transfers a pyrophosphate group
nucleosides and free bases in the intestine, but the prod-
from ATP to C-1 of ribose-5-phosphate, forming PRPP.
ucts are poorly absorbed. Especially the absorbed
PRPP is the activated form of ribose for nucleotide
purine bases are extensively degraded in the intestinal
synthesis. The next enzyme, PRPP amidotransferase,
mucosa. Therefore humans depend on the endogenous
replaces the pyrophosphate group of PRPP with a nitro-
synthesis of purines and pyrimidines. This chapter dis-
gen from the side chain of glutamine. This reaction is the
cusses the pathways for the synthesis and degradation
committed step of purine biosynthesis. In the following
of the nucleotides.
reactions, the purine ring is constructed from simple
building blocks:

CO2 Glycine
NH2 O
Aspartate C
C N C N 6 N
6
N1 5 C 7 HN C N1 5 C 7
2 4
8 CH CH 8C
HC 3 C 9
C C
N N H2N N N C 2 4 C 9
H H 3
N Formyl-THF
N
Adenine Guanine Formyl-THF
Glutamine
NH2 O O

C C C CH3 where THF ¼ tetrahydrofolate. The first nucleotide

N3 4
CH HN CH HN C
5
formed in the pathway is inosine monophosphate (IMP),
2 6
C 1 CH C CH C CH which contains the base hypoxanthine. IMP is a branch
O N O N O N
H H H
point in the synthesis of AMP and GMP (Fig. 28.3). These
Cytosine Uracil Thymine
nucleoside monophosphates are in equilibrium with their
corresponding diphosphates and triphosphates through
Figure 28.1 Structures of the purine and pyrimidine bases. kinase reactions.

471
472 METABOLISM

H2C—O— P H2C—O — P H2C—O— P

O ATP AMP O H2O, PPi, O NH2
glutamine glutamate

OH O— P — P

OH OH OH OH OH OH
Ribose-5-phosphate Phosphoribosyl pyrophosphate Phosphoribosylamine
(PRPP)
ATP,
glycine

ADP, Pi

+
NH 2 H+ H2O, NH NH3
CH2 CHO ADP, Pi, ATP CH2 CHO H+, CH2
glutamate glutamine THF Formyl-THF
C C C
HN NH O NH O NH

P —Ribose P —Ribose P —Ribose

H2O O
–OOC H
N N –OOC —C —N — C
N
HC C ATP, Pi, H C
CH CO2 H+ CH aspartate ADP CH2 CH
C C C
N N COO– N
H2N H2N H2N
P —Ribose P —Ribose P —Ribose

Fumarate

O O O

C N C N C N
H2O THF Formyl-THF H N
HN C H2N C 2 C
CH CH CH
HC C OHC C C
N N N N
N H2N
H
P —Ribose P —Ribose P —Ribose
Inosine monophosphate
(IMP)

Figure 28.2 De novo pathway of purine biosynthesis. THF, Tetrahydrofolate.

As expected, purine synthesis is regulated by feed- base and ribose-1-phosphate by purine nucleoside phos-
back inhibition (Fig. 28.4). The first two enzymes of phorylase. Adenosine is a poor substrate of the nucleo-
the pathway, PRPP synthetase and PRPP amidotrans- side phosphorylase. Therefore it is deaminated to
ferase, are inhibited by the purine nucleotides. In inosine first (Fig. 28.5).
addition, the reactions leading from IMP to AMP and Uric acid is the end product of purine degradation in
GMP are feedback inhibited by the end products. humans. It is synthesized by xanthine oxidase via hypo-
xanthine and xanthine (reaction 6 in Figure 28.5).
This enzyme contains flavin adenine dinucleotide
PURINES ARE DEGRADED TO URIC ACID
(FAD), nonheme iron, and molybdenum. Like other
The degradation of purine nucleotides starts with the nonmitochondrial flavoproteins, it regenerates its FAD
hydrolytic removal of phosphate from the nucleotides. by transferring hydrogen from FADH2 to molecular
The nucleosides thus formed are then cleaved into free oxygen, forming hydrogen peroxide.
 The Metabolism of Purines and Pyrimidines 473

–OOC—CH —CH —COO–

2

NH 2 H+ O O
Pi, GTP,
C N GDP aspartate C N NAD+, H2O NADH, H+ C N
N C HN C HN C
CH CH CH
HC C HC C C C
N N N N O N N
H
P —Ribose P —Ribose P —Ribose
Adenylosuccinate Inosine monophosphate Xanthosine monophosphate
(IMP) (XMP)
ATP, glutamine, H2O
Fumarate AMP, PPi, glutamate, H+

NH2 O

C N C N
N C HN C
CH CH
HC C C C
N N N N
H2N

P —Ribose P —Ribose
Adenosine monophosphate Guanosine monophosphate
(AMP) (GMP)

Figure 28.3 Synthesis of AMP and GMP from IMP.

HGPRT
Ribose 5-phosphate Hypoxanthine + PRPP IMP + PPi
– – Guanine + PRPP HGPRT GMP + PPi
APRT
Adenine + PRPP AMP + PPi
PRPP
– –
The salvage reactions are the only source of purine
– nucleotides for tissues that cannot synthesize the
Phosphoribosylamine
nucleotides de novo. HGPRT is quantitatively by far
the more important salvage enzyme because most of
the adenine is released as hypoxanthine. HGPRT is
IMP competitively inhibited by IMP and GMP, whereas
– – APRT is inhibited by AMP.
ATPGADPGAMP GMPGGDPGGTP

Figure 28.4 Feedback inhibition of de novo purine PYRIMIDINES ARE SYNTHESIZED FROM
biosynthesis by nucleotides. ADP, Adenosine diphosphate; CARBAMOYL PHOSPHATE AND ASPARTATE
AMP, adenosine monophosphate; ATP, adenosine Most proliferating cells synthesize pyrimidines de novo,
triphosphate; GDP, guanosine diphosphate; GMP, guanosine
whereas quiescent cells synthesize pyrimidine nucleo-
monophosphate; GTP, guanosine triphosphate; IMP,
tides from imported bases. Most cancer cells have
Inosine monophosphate; PRPP, 5-phosphoribosyl-1-
pyrophosphate. highly active de novo pyrimidine synthesis.
Unlike the purine ring, the pyrimidine ring is synthe-
sized before the ribose is added (Fig. 28.6). The path-
way starts with carbamoyl phosphate and aspartate,
FREE PURINE BASES CAN BE SALVAGED
and orotic acid is formed as the first pyrimidine. Orotic
As an alternative to degradation, the free bases can be acid is processed to the uridine nucleotides, which are
recycled into the nucleotide pool. This requires the the precursors of the cytidine nucleotides. The enzymes
PRPP-dependent salvage enzymes hypoxanthine-guanine of the pathway are cytosolic except for dihydroorotate
phosphoribosyltransferase (HGPRT) and adenine phos- dehydrogenase (reaction 4 in Figure 28.6), which is on
phoribosyl transferase (APRT): the outer surface of the inner mitochondrial membrane.
474 METABOLISM

ATPGADPGAMP IMP GMPG GDPGGTP

2
1 H2O NH3 1 1
Pi Pi PPi Pi PPi
PPi Adenosine Inosine Guanosine
3 8 8
7 Pi H O NH3 Pi Pi
2
4 4 PRPP 4 PRPP
PRPP Ribose 1— P Ribose 1— P Ribose 1— P

NH2 O O

C N C N C N
N C HN C HN C
CH CH CH
HC C HC C C C
N N N N N N
H2N
H H H
Adenine Hypoxanthine Guanine

H2O, O2
(FAD) 6 5 H2O
H2O2
NH3
O O
H
C N C
6 N
HN C HN C
CH C O
C C C C
O N N H2O, O2 (FAD) H2O2 O N N
H H H H
Xanthine Uric acid

Figure 28.5 Degradation of purine nucleotides to uric acid, and the salvage of purine bases. 1 , 50 -Nucleotidase;
2 , AMP deaminase; 3 , adenosine deaminase; 4 , purine nucleoside phosphorylase; 5 , guanine deaminase; 6 , xanthine
oxidase; 7 , adenine phosphoribosyltransferase; 8 , hypoxanthine-guanine phosphoribosyltransferase. IMP, Inosine
monophosphate; PRPP, 5-phosphoribosyl-1-pyrophosphate.

The carbamoyl phosphate for pyrimidine biosynthe- reduction of ribose to 2-deoxyribose and methylation
sis is synthesized by the cytoplasmic carbamoyl phos- of uracil to thymine.
phate synthetase II. Unlike the mitochondrial enzyme, Ribonucleotide reductase reduces the ribose residue
which makes carbamoyl phosphate for the urea cycle, in all four ribonucleoside diphosphates (Fig. 28.8, A).
the cytoplasmic enzyme uses the side chain of glutamine Its level rises immediately preceding the S phase of the
as a source of nitrogen. cell cycle. It is also subject to intricate allosteric control.
The first three enzymes of the pathway, including carba- dATP is a negative effector for all reactions, whereas
moyl phosphate synthetase II, aspartate transcarbamoy- other nucleotides modulate the substrate specificity
lase, and dihydroorotate dehydrogenase (enzymes 1, 2, and thereby guarantee a balanced production of the
and 3 in Fig. 28.6), are formed by different domains of a four deoxyribonucleotides.
single large polypeptide. Both this multienzyme complex Thymine is synthesized by thymidylate synthase. In
and the CTP synthetase are feedback inhibited by CTP. this reaction, the methylene group of tetrahydrofolate
An inhibitor of dihydroorotate dehydrogenase, leflu- is reduced to a methyl group during its transfer to
nomide, blocks pyrimidine biosynthesis. It has been dUMP, and tetrahydrofolate is oxidized to dihydrofo-
used as an immunomodulatory drug for the treatment late (see Fig. 28.8, B). The active coenzyme form, tetra-
of rheumatoid arthritis and psoriatic arthritis. hydrofolate, has to be regenerated by dihydrofolate
The pyrimidines are degraded to water-soluble prod- reductase (see Fig. 28.8, C).
ucts that are either excreted as such or oxidized to carbon
dioxide and water (Fig. 28.7). MANY ANTINEOPLASTIC DRUGS INHIBIT
NUCLEOTIDE METABOLISM
DNA SYNTHESIS REQUIRES
The development of drugs with selective toxicity for
DEOXYRIBONUCLEOTIDES
cancer cells is difficult because cancer cells are too sim-
The synthesis of 2-deoxyribonucleotides from the ilar to normal cells. Therefore drugs that kill cancer
corresponding ribonucleotides requires two reactions: cells are likely to kill normal cells as well. Cancer cells
 The Metabolism of Purines and Pyrimidines 475

CO2 + Glutamine + 2 ATP

2 H2O
1
O
Glutamate, 2 ADP, Pi, 3 H+
–OOC –OOC
H+ H2O C
Pi, H+ H N CH2 HN CH2
NH2 CH 2
2
+
C CH 2 C C—COO– 3 C C—COO–
O O— P O N O N
+
H3N COO– H H
H H
Carbamoyl Aspartate N-Carbamoyl- L-Dihydroorotate
phosphate aspartate
NAD+
4
NADH, H+

O O O
H+,
C CO2 H+ C PPi PRPP C
HN CH HN CH HN CH

C CH 6 C C 5 C C
O N O N COO– O N COO–
H
P —Ribose P —Ribose Orotate
Uridine monophosphate Orotidine monophosphate
(UMP) (OMP)

NH2
UDP ATP, H2O, ADP, Pi, 2 H+,
glutamine glutamate C
N CH
UTP
7 C CH
O N

P — P — P —Ribose
Cytidine triphosphate
(CTP)

Figure 28.6 Biosynthesis of pyrimidine nucleotides. 1 , Carbamoyl phosphate synthetase II; 2 , aspartate
transcarbamoylase; 3 , dihydroorotase; 4 , dihydroorotate dehydrogenase; 5 , orotate phosphoribosyltransferase;
6 , orotidylate decarboxylase; 7 , CTP synthetase. PRPP, 5-Phosphoribosyl-1-pyrophosphate.

CLINICAL EXAMPLE 28.1: Hereditary Orotic Aciduria

Deficiencies of orotate phosphoribosyltransferase and/or by pyrimidine deficiency. The patients are pyrimidine
orotidylate decarboxylase (reactions 5 and 6 in auxotrophs who can be treated quite effectively with
Figure 28.6), which are two enzymatic activities of a large doses of orally administered uridine.
single protein, lead to hereditary orotic aciduria Orotic aciduria is also seen in some urea cycle enzyme
(Table 28.1). This rare condition is characterized by deficiencies, in which carbamoyl phosphate accumulates
megaloblastic anemia, a crystalline sediment of orotic in liver mitochondria. Some of this leaks into the
acid in the urine, and poor growth. The important clinical cytoplasm, where it is converted to orotic acid (see
signs are caused not by orotic acid accumulation but Clinical Example 26.1).

Table 28.1 Inherited Disorders of Purine and Pyrimidine Metabolism

Disease Enzyme Deficiency Signs and Symptoms

Adenosine deaminase deficiency Adenosine deaminase Severe combined immunodeficiency

Purine nucleoside phosphorylase deficiency Purine nucleoside phosphorylase Immunodeficiency with T-cell defect
Familial orotic aciduria Orotate phosphoribosyltransferase Accumulation of orotic acid in blood and
urine, failure to thrive
Lesch-Nyhan syndrome Hypoxanthine-guanine Mental retardation with self-mutilation,
phosphoribosyltransferase hyperuricemia
476 METABOLISM

O O

C C
Cytidine
HN CF HN CH

Deamination C CH C CH
NH3
O N O N
Uridine 2-Deoxythymidine
Pi P -D-Ribose P -D-Ribose
Nucleoside Pi
Ribose- phosphorylase
1-phosphate 5-Fluorodeoxyuridine Deoxyuridine
2-Deoxyribose-
monophosphate monophosphate
1-phosphate
O O (dUMP)

C C CH3
HN CH HN C This product binds tightly to thymidylate synthase as
a structural analog of its natural substrate dUMP.
C CH C CH
O N O N
Eventually it reacts covalently with the enzyme,
H H resulting in irreversible inhibition. 5-Fluorouracil is
Uracil Thymine also incorporated into RNA in place of uracil, and
NADPH, H+ NADPH, H+ this contributes to its antineoplastic activity.
Reduction
NADP+ NADP+ 2. Antifolates are best exemplified by amethopterin
(methotrexate):
H2O Ring H2O N N
H2N
cleavage C C CH
H+ H+

O O N C C
C N CH2 O COO–
–O—C –O—C CH3
H2N H2N CH NH2 N C N CH
CH2

C CH2 C CH2 CH3 H (CH2)2

O N O N
H H COO–
H2O, H+ H2O, H+ Amethopterin
Hydrolysis (methotrexate)
NH3, CO2 NH3, CO2
H2N N N

O O C C CH

–O—C –O—C CH3 N C C

CH2 CH C N CH2 O COO–

CH2 CH2 OH N C N CH
H3+N H3+N
H H (CH2)2
β -Alanine β -Aminoisobutyrate
COO–
Figure 28.7 Degradation of pyrimidines.
Folic acid

do, however, have a higher mitotic rate than normal Methotrexate inhibits dihydrofolate reductase com-
cells. Therefore they have a higher requirement for petitively, thereby depleting the cell of tetrahydrofo-
DNA synthesis and nucleotide synthesis. With this in late. Thymidylate synthase is the only important
mind, drugs have been developed as antagonists of enzyme that converts a tetrahydrofolate coenzyme to
nucleotide synthesis: dihydrofolate. Therefore rapidly dividing cells, with
their high activity of this enzyme, are most vulnerable
1. Structural analogs of bases or nucleosides act
to methotrexate.
either as inhibitors of nucleotide biosynthesis or
through their incorporation into DNA or RNA. These anticancer drugs cause collateral damage to rap-
5-Fluorouracil is used to treat cancers of the colon, idly dividing cells in bone marrow, intestinal mucosa,
pancreas, stomach, esophagus, and breast. It is and hair bulbs. Therefore bone marrow depression,
processed to fluorodeoxyuridine monophosphate diarrhea, and hair loss are common side effects of can-
in the body: cer chemotherapy.
 The Metabolism of Purines and Pyrimidines 477

Figure 28.8 Synthesis of

Base Base
2-deoxyribonucleotides, the
P—P O CH2 O NADPH, H+ NADP+, H2O P — P O CH2 O
precursors for DNA synthesis.
DHF, Dihydrofolate; dTMP,
H H Ribonucleotide reductase H H
H H H H deoxythymidine monophosphate;
dUMP, deoxyuridine
OH OH OH H monophosphate; THF,
tetrahydrofolate.
Ribonucleoside 2-Deoxyribonucleoside
diphosphate diphosphate
A

O O

C Methylene-THF DHF C
HN CH HN C CH3

C CH Thymidylate synthase C CH
O N O N

P -D-Ribose P -D-Ribose
B dUMP dTMP

NADPH, H+ NADP+
Dihydrofolate Tetrahydrofolate
(DHF) Dihydrofolate (THF)
C reductase

CLINICAL EXAMPLE 28.2: Severe Combined Immunodeficiency

Deficiency of adenosine deaminase (ADA) leads to an precursors for DNA synthesis. We do not know why
autosomal recessive form of severe combined lymphocytes are more sensitive than other cells to this
immunodeficiency (SCID), a type of disease with metabolic defect.
combined B-cell and T-cell defects. ADA deaminates Enzyme replacement therapy with injected bovine
adenosine to inosine and deoxyadenosine to adenosine deaminase is possible, but its usefulness is
deoxyinosine (Fig. 28.9). limited by high cost and by immunological reactions to
Both adenosine and deoxyadenosine can also be the injected enzyme. Gene therapy is more promising.
phosphorylated to the corresponding nucleoside For gene therapy, hematopoietic stem cells from the
monophosphate. However, whereas AMP can be patient are treated in vitro with an ADA-containing
deaminated to IMP by AMP deaminase, dAMP has no retroviral vector. After partial destruction of the patient’s
alternative route of degradation. It accumulates, together bone marrow by chemotherapy, the engineered cells are
with its diphosphate and triphosphate derivatives. dATP reinserted in the patient. Unencumbered by ADA
is thought to cause the immunodeficiency by inhibiting deficiency, the engineered cells can take over the
ribonucleotide reductase, thus depriving the cell of ecosystem from the unmodified lymphocytes.

NH3 H O Figure 28.9 Role of adenosine

2
deaminase in the metabolism of
IMP AMPGADPGATP dAMPGdADPGdATP adenine nucleotides. The
AMP deaminase
H2O ADP H2O
ribonucleotides can be
ADP
catabolized by AMP deaminase,
Pi ATP ATP Pi but the deoxyribonucleotides
accumulate in adenosine
Adenosine 2-Deoxyadenosine
deaminase deficiency. dADP,
H2O H2O
Adenosine Adenosine deoxy-ADP; dAMP, deoxy-AMP;
deaminase deaminase dATP, deoxy-ATP; IMP, inosine
NH3 NH3
monophosphate; Pi, inorganic
Inosine 2-Deoxyinosine
phosphate.
478 METABOLISM

URIC ACID HAS LIMITED WATER SOLUBILITY over time. On random sampling, between 2% and
18% of healthy people have uric acid levels above the
All purines are catabolized to uric acid. Although not
solubility limit of 7 mg/dl, and 10% to 20% of gouty
very toxic, uric acid has a serious problem: low water sol-
patients have levels below this limit at the time of their
ubility. It can form damaging crystals both in the urine and
first attack.
in the tissues. Uric acid is a weak acid, with a pK of 5.7,
Synovial fluid analysis from an acutely inflamed joint
and its water solubility depends on its ionization state
shows needle-shaped optically birefringent crystals of
(Fig. 28.10).
sodium urate, often within polymorphonuclear leuko-
The protonated form usually is less soluble than the
cytes. These cells phagocytize sodium urate crystals,
deprotonated form. In urine of pH 5.0, uric acid
but the razor-sharp, undigestible crystals damage their
becomes insoluble at concentrations above 0.9 mmol/L
lysosomes and thereby kill the cell.
(15 mg/dl). Uric acid stones can form in the collecting
The cause of primary hyperuricemia usually is un-
ducts, where the urine becomes concentrated and acid-
known, but secondary hyperuricemia is caused by an
ified. Between 5% and 10% of all kidney stones consist
underlying disease. It is seen in psoriasis, chronic hemo-
of uric acid.
lytic anemias, pernicious anemia, malignancies, and
In plasma and interstitial fluids, with a pH of 7.3 to
other conditions with increased cell turnover. Radiation
7.4 and a high sodium concentration, sodium urate is
treatment or chemotherapy for neoplastic diseases can
the least soluble form. It tends to precipitate at concen-
cause massive hyperuricemia, with a risk of uric acid
trations above 0.4 mmol/L (7 mg/dl). Most adults have
nephropathy and renal failure.
serum urate levels between 3 and 7 mg/dl. Therefore
Uric acid production is also increased in metabolic
even a moderate rise of the serum urate concentration
disorders in which the activity of the pentose phosphate
will exceed the limit of solubility. The average uric acid
pathway is increased. In type I glycogen storage disease
level is higher in men than in women by about 1 mg/dl
(von Gierke disease; see Chapter 22), for example,
and rises with increasing age.
accumulating glucose-6-phosphate is converted into
ribose-5-phosphate by the pentose phosphate pathway.
HYPERURICEMIA CAUSES GOUT
Ribose-5-phosphate feeds into purine nucleotide bio-
A serum uric acid level above the limit of solubility synthesis, and the excess nucleotides are degraded to
(hyperuricemia) can lead to the formation of sodium uric acid. Purines cannot be stored in the body; there-
urate crystals in the tissues. Focal deposits of sodium fore, any increase in the rate of their de novo synthesis
urate in subcutaneous tissues, known as tophi, are has to be matched by an increased rate of degradation
asymptomatic, but sodium urate crystals in the joints to uric acid.
trigger the inflammatory response of gouty arthritis. Primary hyperuricemia is caused by overproduction of
Gout is a common disease. In Britain, the prevalence uric acid, impairment of its renal excretion, or both.
of gout is 1.4% in all men and 7% in men over the Urinary excretion in excess of 600 mg/day is evidence of
age of 65 years. The prevalence is nearly four times uric acid overproduction. From 15% to 25% of patients
higher in men than in women. with primary gout are overproducers; the other 75% to
Gouty arthritis takes the form of acute attacks of 85% of patients have reduced renal clearance of uric acid.
joint pain and inflammation, separated by asymptom- Renal excretion is complex. Uric acid is first reabsorbed
atic intervals. The disease has a predilection for small and then actively secreted in the tubular system.
peripheral joints, and the metatarsophalangeal joint of Most animals other than the higher primates do not
the big toe is initially affected in about half of the develop gout because they degrade uric acid to water-
patients. This is because the solubility of sodium urate soluble products. Why do humans use uric acid as the
is temperature dependent, and crystals form in the cold- end product of purine metabolism, and why is our uric
est parts of the body first. acid level so high that we are teetering on the brink of
Any sustained hyperuricemia is likely to cause gouty gout? One possible reason is that uric acid is an antiox-
arthritis, but uric acid levels are somewhat variable idant that scavenges hydroxyl radicals, superoxide

Figure 28.10 Water solubility

of uric acid depends on its O O O
H
ionization state. Its pK value C C N C N
N
is 5.7. HN C G HN C G HN C
C O C OH C O– + H+
C C C C C C
O N N O N N O N N
H H H H H H

Uric acid Uric acid Urate

(keto form) (enol form)
 The Metabolism of Purines and Pyrimidines 479

radicals, singlet oxygen, and other aggressive oxygen

The brain has a very low capacity for de novo purine
derivatives. It contributes to the body’s defenses against
biosynthesis; therefore, it depends on the salvage
oxidative damage.
enzyme for its purine nucleotides. Although the cause
of the neurological symptoms is not known, existing data
ABNORMALITIES OF PURINE-METABOLIZING indicate that there is a loss of the neurotransmitter
ENZYMES CAN CAUSE GOUT dopamine in specific neurons in the basal ganglia region
Abnormalities of two enzymes have been identified in a of the brain.
minority of patients with uric acid overproduction:
1. Overactivity of PRPP synthetase increases de novo
purine biosynthesis, and purine breakdown must rise GOUT CAN BE TREATED WITH DRUGS
in proportion. Elevated levels of IMP, GMP, and
The immediate aim in the treatment of gout is the allevi-
AMP from increased de novo biosynthesis inhibit
ation of pain and inflammation, but long-term treatment
the salvage enzymes and thereby favor uric acid for-
is aimed at reducing the serum uric acid level. The most
mation over recycling. Hyperuricemia in individuals
important drug treatments are as follows:
with an overactive PRPP synthetase shows that this
enzyme is normally rate limiting for de novo purine 1. Antiinflammatory drugs. Colchicine is the classic treat-
synthesis. The rate of the amidotransferase reaction ment of the acute attack. It is not very effective in other
depends largely on the concentration of its rate-limiting forms of arthritis; therefore, it can be used for the
substrate PRPP. differential diagnosis of gout. This approach, in which
2. Reduced activity of the salvage enzyme HGPRT the response to treatment confirms (or refutes) a prelim-
causes hyperuricemia because the substrates of the inary diagnosis, is called a diagnosis ex juvantibus.
deficient enzyme accumulate, whereas the levels of Because of gastrointestinal side effects, however,
its products are reduced: colchicine has been largely replaced by indomethacin,
ibuprofen, and other nonsteroidal antiinflammatory
l The cellular levels of the free bases are increased,
drugs.
thus providing more substrate for uric acid synthesis.
2. Uricosuric agents increase the renal excretion of uric
l The cellular PRPP level is increased because of
acid. Probenecid is an example.
decreased consumption in the salvage reactions,
3. Inhibition of xanthine oxidase is possible with allopuri-
and more substrate is available for the PRPP ami-
nol, a purine analog that is oxidized to alloxanthine by
dotransferase of the de novo pathway.
xanthine oxidase. Alloxanthine remains bound to the
l The cellular concentrations of the nucleotides are
enzyme as a competitive inhibitor. As a result, the
reduced. This disinhibits the regulated enzymes
patient excretes a mix of uric acid, xanthine, and hypo-
of the de novo pathway. The result is an increased
xanthine that is more soluble than uric acid alone.
rate of de novo synthesis, balanced by an equally
Hypoxanthine and xanthine are turned into IMP and
increased rate of uric acid formation.
xanthosine monophosphate (XMP), respectively, by
Both PRPP synthetase and HGPRT are encoded by HGPRT. These salvage reactions consume PRPP and
genes on the X chromosome, and the enzyme abnor- produce nucleotides that feedback-inhibit the regu-
malities are expressed as X-linked recessive traits. lated enzymes of the de novo pathway, thereby reduc-
ing de novo purine biosynthesis. A new xanthine
oxidase inhibitor, febuxostat, acts noncompetitively
CLINICAL EXAMPLE 28.3: Lesch-Nyhan
by blocking the channel leading to the active site.
Syndrome
Dietary manipulations are less effective. Dietary pur-
Partial deficiencies of HGPRT cause only hyperuricemia,
ines are degraded to uric acid in the intestinal mucosa.
but complete deficiency results in Lesch-Nyhan
Although much of this uric acid ends up in the stools,
syndrome. This rare X-linked recessive disorder is
in which it is degraded by intestinal bacteria, the con-
characterized by dystonia, choreoathetosis, spasticity,
sumption of 4 g of yeast RNA per day raises the blood
mental retardation, and bizarre self-mutilating behavior.
urate to levels typical for gout. On the other hand, elim-
If unrestrained, the patients chew off their lips and
inating all purines from a typical diet would reduce the
fingers or jam their hands in the spokes of their
serum urate level by only 1 mg/dl.
wheelchairs. The presence of uric acid crystals in the
Alcohol should be avoided because of associated
urine is an early sign of the disease, and many patients
dehydration and because the increased lactate level dur-
eventually die of uric acid nephropathy.
ing alcohol intoxication can impair the renal excretion
The brain disorder, however, is not caused by uric
of uric acid. Acute attacks of gouty arthritis are often
acid overload but by the deficiency of purine nucleotides.
triggered by an alcoholic binge.
480 METABOLISM

SUMMARY Further Reading

Aiuti A, Cattaneo F, Galimberti S, et al: Gene therapy for
Nearly all purines and pyrimidines in the human body
immunodeficiency due to adenosine deaminase deficiency,
are derived from endogenous synthesis. The heterocy- N Engl J Med 360:447–458, 2009.
clic ring systems are assembled from simple precursors, Alcorn N, Saunders S, Madhok R: Benefit-risk assessment of
whereas the ribose portion of the nucleotides comes leflunomide: an appraisal of leflunomide in rheumatoid
from PRPP, the activated form of ribose-5-phosphate. arthritits 10 years after licensing, Drug Saf 32:1123–1134,
The first reactions of the biosynthetic pathways are 2009.
feedback inhibited by the nucleotides. An S, Kumar R, Sheets ED, et al: Reversible compartmentali-
The ribonucleotides are the precursors of the corres- zation of de novo purine biosynthetic complexes in living
ponding 2-deoxyribonucleotides. Rapidly dividing cells cells, Science 320:103–106, 2008.
depend on a high rate of nucleotide biosynthesis; there- Evans DR, Guy HI: Mammalian pyrimidine biosynthesis:
fore, inhibitors of nucleotide biosynthesis can be used fresh insights into an ancient pathway, J Biol Chem
279:33035–33038, 2004.
for cancer chemotherapy.
Jinnah HA, Ceballos-Picot I, Torres RJ, et al: Attenuated var-
Purine nucleotides are catabolized to the free bases, iants of Lesch-Nyhan disease, Brain 133:671–689, 2010.
which are either oxidized to the excretory product uric Riches PL, Wright AF, Ralston SH: Recent insights into the
acid or recycled to the corresponding nucleotides in pathogenesis of hyperuricaemia and gout, Hum Mol Genet
PRPP-dependent salvage reactions. Uric acid is poorly 18:R177–R184, 2009.
soluble in water. Therefore it can cause kidney stones, Saag KG, Choi H: Epidemiology, risk factors, and lifestyle
and it causes gout when crystals of sodium urate form modifications for gout, Arthritis Res Ther 8(Suppl 1):S2,
in the joints. 2006.
Segal NH, Saltz LB: Evolving treatment of advanced colon
cancer, Annu Rev Med 60:207–219, 2009.

QUESTIONS

1. Enzyme abnormalities that can lead to 2. A coenzyme form of tetrahydrofolate is

hyperuricemia and gout include required for
A. Reduced activity of PRPP synthetase A. The de novo synthesis of pyrimidines
B. Reduced activity of xanthine oxidase B. The synthesis of thymine-containing nucleotides
C. Reduced activity of hypoxanthine-guanine from uracil-containing nucleotides
phosphoribosyltransferase C. The synthesis of xanthine from purine nucleotides
D. Reduced activity of PRPP aminotransferase D. The cleavage of the purine ring in uric acid by an
E. Reduced activity of dihydroorotate enzyme in the human liver
dehydrogenase E. The reduction of ribonucleotides to
2-deoxyribonucleotides