Salvage Pathway for Nucleotide

Biosynthesis

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Salvage Pathways for Nucleotide Biosynthesis
- The pathways produce nucleotides from preformed purine and
pyrimidine bases and nucleosides
- Endogenous NA breakdown, foreign DNA and RNA, which enters body
through infectious agents breakdown and digestion of dietary NAs are the
sources for preformed bases and nucleosides
- Synthesis of nucleotides from preformed bases and nucleosides saves
considerable cellular energy
- Tissues like erythrocytes, leukocytes and brain lack enzymes of de
novo pathways and hence they entirely depends on salvage pathways for
nucleotide biosynthesis
- Liver supplies free bases and nucleosides to salvage pathways of brain,
erythrocytes and leukocytes
- The salvage pathways helps in recycling of 90% of preformed bases and
nucleosides in the body

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Purine Salvage Pathways

- In the blood, the concentration of guanine and hypoxanthine is higher

than adenine
- Guanine and hypoxanthine are salvaged by hypoxanthine-guanine
phosphoribosyl transferase (HGPRTase) which converts hypoxanthine and
guanine to IMP and GMP respectively by using PRPP as donor of ribose-5-
phosphate
- Adenine is salvaged by adenine phosphoribosyl transferase and converts
adenine to AMP using PRPP as donor of ribose-5-phosphate
- Free guanine is formed from guanosine by removing ribose as ribose-1-
phosphate, catalyzed by purine nucleoside phosphorylase. The enzyme
also acts on deoxyguanosine. Free hypoxanthine is formed from adenosine
via inosine.
- Adenosine deaminase (ADA) converts adenosine to inosine, followed by
release of inosine ribose as ribose-1-phosphate. ADA acts on
deoxyadenosine
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Purine Salvage Pathways

- Purine nucleoside phosphorylase catalyzes latter reaction and

hypoxanthine is the product
- Adenosine nucleosidase catalyzes the formation of adenine from
adenosine by removing ribose as ribose-1-phosphate
- Purine nucleosides are salvaged by distinct kinases which phosphorylates
nucleosides
- Adenosine kinase catalyzes conversion of adenosine to AMP. Deoxy
adenosine and deoxyguanosine are salvaged by deoxy adenosine kinase

Energy aspect of purine salvage pathways

- Only two high-energy bonds are required for the conversion of free bases
to nucleoside mono phosphates (one high energy bond for PRPP formation
and another high energy bond by way of PP hydrolysis)
- One high energy bond is used for the formation of nucleoside
monophosphates from nucleosides
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Summary of Purine Salvage Pathways

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Pyrimidine Salvage Pathways
- Free pyrimidine bases are salvaged by pyrimidine phosphoribosyl
transferase that catalyses conversion of uracil or thymine to UMP and TMP
using PRPP as donor of ribose phosphate

- It also acts on 5-fluorouracil and orotate

- Thymine is salvaged by thymidine phosphorylase. It catalyzes conversion

of thymine to deoxy thymidine by incorporating deoxy ribose

- Pyrimidine nucleosides are salvaged by distinct pyrimidine nucleoside

kinases. They phosphorylate nucleosides using ATP as phosphate donor

- Cytidine and deoxy cytidine are phosphorylated by cytidine kinase and

deoxy cytidine kinase, respectively

- Likewise deoxy thymidine is phosphorylated by thymidine kinase

- Uridine is phosphorylated by uridine kinase

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Pyrimidine Salvage Pathways

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 Medical Importance
Salvage pathways are affected in some diseases or syndromes
1. Immuno deficiency disease
It may be due to
a). Lack of adenosine deaminase (ADA) - leads to accumulation of
deoxy adenosine and increased production of dATP from deoxy
adenosine by salvage pathway
- T-lymphocytes and B-lymphocytes do not mature due to impaired DNA
synthesis
- Condition is usually called as severe combined immunodeficiency
disease (SCIDD)
b). Deficiency of purine nucleoside phosphorylase
- Milder form only T-lymphocyte production is impaired
- Leads to accumulation of deoxy guanosine and dGTP which blocks
conversion of CDP to dCDP by ribonucleotide reductase
- dCTP required for a DNA synthesis is not available and proliferation of
T-cells is impaired 8
 Medical Importance
2. Lesch-Nyhan syndrome
- It is due to lack of HGPRT ase

- Lack of this enzyme leads to accumulation of PRPP and decreased IMP

or GMP levels

- Since PRPP is a positive effector of amidotransferase purine

nucleotide synthesis is promoted

- Low levels of IMP or GMP also promotes purine nucleotide

biosynthesis because feed back inhibition of amidotransferase by
GMP or IMP is decreased

- Symptoms - hyperuricemia, mental retardation, self mutilation and

anaemia
- Uric acid excretion is more in urine 9
Nucleotide metabolism of malarial parasite P. faliciparum
- P. falcipuram synthesizes purines by the salvage pathway and pyrimidines by de
novo pathways

- AMP and GMP are synthesized from IMP which is formed from hypoxanthine by
HGPRTase catalyzed reaction

- In human cells, purines are synthesized by de novo and pyrimidines are

synthesized by either salvage or de novo pathways

- In P. falcipuram first three enzymes of pyrimidine biosynthesis carbamoyl

phosphate synthetase, aspartate transcarbamoylase and dihydrorotase exist as
a separate independent units. In host human cells, they exist as single unit

- Many other parasitic organisms are unable to synthesize purine nucleotides via
de novo pathways. Salvage pathways are main supplies of purine nucleotides in
these parasites
- e.g. in Giardia lamblia causative agent of giardiasis lacks de novo pathways of
purine biosynthesis; Trichomones fetus which causes embryonic death and
infertility in cows depends primarily on a single enzyme of salvage pathway
HGPRTase for its purine needs - Enzymes in salvage pathways potential
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targets of new therapeutic agents
Digestion of Dietary Nucleic Acids

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Degradation of Purine Nucleotides
- Liver is the major organ involved in degradation of purine nucleotides

- Lysosomal enzymes converts nucleic acids to nucleotides

- AMP is converted to IMP by adenylate deaminase

- Nucleotidases convert IMP, AMP and GMP to corresponding nucleosides

namely inosine, adenosine and guanosine

- By the action of adenosine deaminase adenosine is converted to inosine

- Purine nucleoside phosphorylase converts guanosine to guanine and

inosine to hypoxanthine by transferring ribose

- Deamination of guanine by guanase produces xanthine

- Hypoxanthine and xanthine are converted to uric acid by xanthine

oxidase
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Degradation of Purine Nucleotides

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Medical Importance
Fate of uric acid
- Uric acid produced in different tissues diffuses into circulation and
carried to kidneys for elimination
- Daily production of uric acid is about 500-600mg, most of it is removed
by kidney. Daily output is about 0.3-0.5 gm/day on normal diet. The
normal blood uric acid level is below 6 mg/100ml. So one can expect
that impaired renal function may lead to accumulation of uric acid in
blood
Gout
- A common disease associated with excessive purine catabolism

- Characterized by hyperuricemia and excessive excretion of uric acid in

urine

- Excessive uric acid leads to formation and deposition of urate crystals in

joints, cartilage of fingers, big toe and other soft tissues

- Deposition of urate in joints leads to gouty arthritic attacks 14

Hyperuricemia or Gout
- Due to over production of uric acid and Impaired excretion of uric
acid
1. Primary gout is due to excessive formation of purine nucleotides
and their degradation. It is an inherited disease. It occurs due to
(a) Deficiency of HGPRT ase (Lesch-Nyhan syndrome)

(b) Increased PRPP synthase activity

2. Secondary gout can be acquired as well as inherited disease. It

occurs as consequence of other diseases, which cause excessive uric
acid production. They are
(a) Leukemia, polycythemia - Nucleic acid turnover is more in both

these diseases which is responsible for uricemia

(b) Von-Gierke’s disease - In this disease, glucose-6-phosphate

accumulates due to lack of glucose-6-phosphatase, which is

diverted to HMP shunt. As a result more pentose phosphates are
produced. This leads to accumulation of PRPP and increased
purine nucleotide biosynthesis
(c) Increased glutathione reductase activity
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3. Renal gout If the hyperuricemia is due to impaired excretion of uric
acid by kidney then it is called as renal gout. It occurs due to defective
uric acid transport in renal tubules and Glomerulonephritis

Treatment - Drugs used in treatment of gout work by lowering uric acid

production or level
Hypoxanthine analog - Allopurinol is the drug used in the treatment of
gout
- It is a hypoxanthine analog which is substrate for xanthine oxidase and
inhibits xanthine oxidase by binding at active site

- Xanthine oxidase converts allopurinol to alloxanthine by hydroxylation

- It is converted to active drug by xanthine oxidase, which subsequently

inactivate the enzyme and thus uric acid production is decreased

- This type of enzyme inactivation is often referred as suicide inhibition

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 Conversion of allopurinol to alloxanthine by xanthine
oxidase

Sulfin pyrazone and proben acid

- These drugs lower uric acid level by increasing its excretion by kidney
- They interfers with tubular reabsorption of uric acid. They are uricosuric
drugs
Xanthinuria
- An inherited disease and characterized by hypouricemia and increased
excretion of hypoxanthine and xanthine in urine
- Hepatic or intestinal xanthine oxidase is deficient. In severe cases,
xanthine lithiasis may occur due to deposition of xanthine crystals in 17
kidney
Degradation of Pyrimidine Nucleotides
- Liver is the major organ involved in breakdown of pyrimidine nucleotides

- Pyrimidine nucleotide are degraded to amino acids beta-alanine and beta-amino

isobutyric acid (BAIB) by cleaving pyrimidine ring

- Catabolism of pyrimidine nucleotides proceeds in three phases

- Nucleases convert nucleic acids to pyrimidine nucleotides

1. In the first phase, pyrimidine nucleotides undergoes dephosphorylation,

deamination and glycosidic bond cleavage reactions to yield free bases

2. In the second phase, uracil and thymine undergo sequence of reduction

(unusual in degradative phase) hydration and deamination reactions to yield
beta-alanine and b- amino isobutyric acid

3. In the third phase, beta-alanine and BAIB undergoes transamination and

activation to yield malonyl-CoA and methyl malonyl-CoA

- Succinyl-CoA is formed from methyl malonyl-CoA. Malonyl-CoA may be converted

to CO2 ultimately
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Degradation of Pyrimidine Nucleotides

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Medical and Biological Importance
1. beta-Amino isobutyric aciduria
- It is familial disease due to deficiency of transaminase, which converts
BAIB to methylmalonic semialdehyde

- This leads to accumulation of BAIB and its increased excretion in urine

2. beta-amino isobutyric aciduria also occurs in leukemia and

radiotherapy

3. beta-amino isobutyric acid excretion in urine depends on turnover of

nucleic acids

4. beta-alanine may be used for the synthesis of coenzyme A and

carnosine

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