NUCLEIC ACIDS

&
METABOLISM

Okunowo O Wahab, Ph.D.

Biochemistry Department
College of Medicine
University of Lagos
E-mail: modelprof@yahoo.com
 Course Outline
 Nucleotide & Nucleic Acid Nomenclature

 Importance of Nucleotides to Life

 Purine Synthesis

i. De Novo Purine Nucleotide Synthesis

ii. Salvage Pathway For Purines

 Degradation of Purine Nucleotides

 Disorders of Purine Catabolism

 Biosynthesis of Pyrimidine Nucleotides

 Disorder of Pyimidine Metabolism

 Degradation of Pyrimidine Nucleotides

 Enzymes Involved in the Digestion of Polynucleotides in the G.I.T

Introduction
 Biomolecules found in the nucleus and
other parts of a cell includes phosphoric
acids and are made up of:

 Purine or pyrimidine base.

 Sugar moiety (ribose or deoxy ribose)

 Phosphate group (Mono,di or tri phosphate)

 Nucleotide & Nucleic Acid Nomenclature
 Purine: Adenine &
Guanine

 Pyrimidine: Thymine,
Cytosine& Uracil

 Nucleoside = Base + sugar

 Nucleotide = Base + sugar

+ phosphate

 Nucleotide = Nucleoside +
phosphate

 DNA & RNA contain the

same purine bases: A&G

 DNA & RNA contain the

pyrimidine Cytosine, but
differ in the second
pyrimidine base

 DNA contain thymine (T)

whereas RNA contain
Uracil (U).
Examples of nucleotides

 DNA & RNA contain the same purine bases: Adenine and
Guanine.
 DNA & RNA contain the pyrimidine Cytosine, but differ in the
second pyrimidine base: DNA contain thymine (T) whereas
RNA contain Uracil (U).
Importance
 Storage & transmission of genetic information from
generation to generation

 Precursor for DNA, RNA & Protein synthesis

 Carries of activated intermediates in the synthesis

of carbohydrate (UDP-glucose), lipids & proteins.

 For conservation & utilization of energy (ADP-ATP)

released during cellular metabolism

 As essential part of coenzymes needed in oxido-

reduction reactions (FAD, NAD, NADP)
PURINE SYNTHESIS
 The atoms of the purine
ring are contributed by a
number of compounds,
including CO2 which is
synthesized in humans
from various metabolic
pathways including TCA
cycle.

 The amino acids are non-

essential (aspartic acid,
glycine, and glutamine)
and can by synthesized
in the body.

 The N10-formyl THF can

be synthesized in
microbes from the
precursor molecules
shown in subsequent
slide.
Synthesis of N -formyl THF 10

This is a major inhibitory step in the series of reactions leading to purine

synthesis. This is so because Sulphonamide or sulfa drugs (PABA analog) and
Methotrexate (folic acid analog) inhibits the enzymes Dihydropteroate synthetase
and Dihyrofolate reductase respectively in THF synthesis. Once THF cannot be
synthesized by the intestinal microflora, its role in the synthesis of purine cannot
be ascertained.
DE NOVO PURINE NUCLEOTIDE SYNTHESIS

 PRPP Synthase is inhibited by purine nucleosides but

activated by Pi

 PRPP generated also participates in the synthesis of

pyrimidines and in the salvage reactions of purines
and pyrimidines

 The next enzyme in the next reaction; glutamine

phosphoribosyl amidotransferase is inhibited by AMP,
GMP, IMP by a feedback mechanism.
The purine ring is constructed by a series of reactions that
add the donated carbons and nitrogens to a preformed ribose
5-phosphate. Ultimately, Inosine monophosphate is formed.
This is then converted to AMP & GMP as shown below.

Biosynthesis of AMP and GMP from IMP with feedback inhibition

IMP, AMP, GMP can be converted to their di or triphophate for example

AMP + ATP Adenylate kinase 2ADP
GMP + ATP Guanylate kinase GDP + ADP
 SALVAGE PATHWAY FOR PURINES

 Purines that result from 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 body.

 This is referred to as “Salvage

pathway” for purines.

 Two enzymes are involved.

 Adenosine phosphoribosyl
transferase (APRT) and hyoxanthine
guanine phosphoribosyl transferase
(HGPRT).

 Both enzymes utilizes PRPP as the

source of the ribose 5-phosphate
group. The release of pyrophosphate
(PP) makes these reactions
irreversible.

 A deficiency of HGPRT causes

Lesch-Nyhan Syndrome
 DEGRADATION OF PURINE NUCLEOTIDES
 The end product of purine
catabolism in humans is uric
acid. In other primates;
allantoin, other mammals
further degrade the uric acid to
urea or even ammonia.

1. Amino group is removed from

AMP to produce IMP, or from
Adenine to produce Inosine.

2. IMP & GMP are converted into

their respective nucleoside
forms inosine & guanosine by
the action of 51-nucleotidase.

3. Purine nucleoside
phosphorylase converts
inosine & guanosine into their
respective purine bases;
hypoxanthine & xanthine

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. Uric acid
is excreted in the urine.
 DISORDERS OF PURINE CATABOLISM
 ADENOSINE DEAMINASE DEFICIENCY:
1. Causes severe combined immunodeficiency involving T-cell & B-cell dysfunction.
This is because there is accumulation of deoxyadenosine which increases
adenosylhomocysteine, this substance is toxic to immature lymphocytes resulting in
immunocompromised immune system.

2. Extremely large buildups of dATP in red cells (this dATP inhibits ribonucleotide
reductase, and therefore DNA synthesis).

3. ADA-deficencient children usually die before 2years of age from overwhelming

infection.

 PURINE NUCLEOTIDE PHOSPHORYLASE DEFECIENCY:

1. Causes impairment of T-cell function but no apparent effect on B-cell function.

2. It is characterised with decreased uric acid formation, combined with increased

levels of purine nucleosides & nucleotides.

3. dGTP is the major nucleotide that accumulates in red cells (dGTP is converted to
dATP, an inhibitor of ribonucleotide reductase. dGTP also inhibits the reduction of
UDP & CDP)

 GOUT:
1. Characterised by hyperuricemia, with recurrent attacks of acute arthritic joint
inflamation caused by uric acid crystals deposition.

2. Primary gout (hyperuricemia) is the form of the disease that is attributable to an in-
born error of metabolism, such as overproduction of uric acid.
 GOUT Contd.:
3. Secondary hyperuricemia may be caused by other diseases, for
example, cancer, chronic renal insufficiency, HGPRT deficiency, e.t.c.

4. Treatment with allopurinol which inhibits xanthine oxidase leading to

accumulation of hypoxanthine & xanthine which are more soluble than
uric acid.

 HYPOURICEMIA
1. Hypouricemia and increased excretion of hypoxanthine and xanthine
are associated with xanthine oxidase deficiency due to a genetic
defect or to severe liver damage.

2. Patients with a severe enzyme deficiency may exhibit xanthinuria and

xanthine lithiasis

 LESCH-NYHAN SYNDROME
1. Lesch-Nyhan Syndrome is a inherited disorder associated with a
virtually complete deficiency of hypoxanthine-guanine
phosphoribosyltransferase, and therefore inability to salvage
hypoxanthine or guanine.

2. This results in excessive production of uric acid, plus characteristic

neurological features including self mutilation, involuntary movements,
and mental retardation.

3. Enzyme deficiency results in increased levels of PRPP and decreased

IMP and GMP, causing increased de novo purine synthesis
Biosynthesis of pyrimidine nucleotides

 Unlike purine ring where the ring is constructed on 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 carbon and nitrogen atoms in the pyrimidine ring are
glutamine, CO2, and aspartic acid.

 In mammalian cells the synthesis of carbamoyl phosphate, CPSII is

inhibited by UTP and activated by ATP & PRPP.

 Synthesis of CTP is produced by amination of UTP by CTP synthase.

The biosynthetic pathway
for pyrimidine nucleotides
DISORDER OF PYRIMIDINE METABOLISM
 OROTIC ACIDURIA
1. Orotate phosphoribosyl transferase and OMP
decarboxylase are separate domains of a single
polypeptide.

2. Low activities of orotidine phosphate decarboxylase

and orotate phosphoribosyltrnsferase result in
abnormal growth, megaloblastic anemia, and the
excretion of large amounts of orotate in the urine.

3. Feeding in a diet rich in uridine results in

improvement of the anemia and decrease excretion
of orotate.
DEGRADATION OF PYRIMIDINE NUCLEOTIDES

 Unlike purine rings, human cells can

open and degrade pyrimidine ring to
highly soluble structures such as β -
alanine and β -aminoisobutyrate that
can serve as precursors of acetyl coA &
Succinyl coA respectively.

 Also, pyrimidine can be salvaged and

converted to nucleotides by the enzyme
pyrimidine phosphoribosyltransferase,
which like its counterparts in purine
salvage reactions, utilizes PRPP as the
source of the ribose-P
 ENZYMES INVOLVED IN THE DIGESTION OF
POLYNUCLEOTIDES IN THE G.I.T
 Ribonucleases and
deoxyribonucleases
secreted in
pancreatic juice,
hydrolyse RNA &
DNA primarily to
oligonucleotide.

 This is further
broken down by
phosphodiesterase,
producing
mononucleotides.

 Nucleotidase
removes Phosphate
group hydrolytically.

 Nucleosidase
degrade the
nucleosides to free
bases; purines &
pyrimidines