Purine Biosynthesis

There are two pathways of synthesis of purine nucleotides:

1. De Novo synthesis pathway

2. Salvage pathway

Significance of Purine Synthesis

1. Purines serve as building blocks of nucleic acids.

2. ATP plays an important role in energy transformation.

3. ATP, ADP, and AMP may function as allosteric regulators and

participate in regulation of many metabolic path-ways

4. cGMP are secondary messengers

5. Salvage pathways are used to recover bases and nucleosides that are

formed during degradation of RNA and DNA.

6. In comparison to de novo pathway, salvage pathway is energy-saving

7. In brain and bone marrow tissues salvage pathway is the only pathway

of nucleotide synthesis

De Novo synthesis pathway

Purine synthesis occurs in all tissues. The major site of purine synthesis

is in the liver and, to a limited extent, in the brain.

Overview of the pathway

 The starting material is ribose 5-phosphate, which is

phosphorylated by PRPP synthetase to PRPP (5′-Phosphoribosyl

1′-pyrophosphate) using two phosphates from ATP.

 In the committed step in the process, an α-amino group is then

added to PRPP from glutamine to form 5-phosphoribosylamine.

This reaction is catalyzed by glutamine PRPP amidinotransferase.

 PRPP amidotransferase is regulated partly by GMP and partly by

AMP. The presence of either of these can reduce the enzyme’s

activity. Only when both are present is the enzyme fully

inactivated.

 A series of nine reactions results in the formation of IMP (Inosine

5′-monophosphate). These reactions include

 adding glycine

 adding carbon (from N 10-formyltetrahydrofolate)

 adding amine (from glutamine)

 closing of the first ring, addition of carboxyl (from CO2CO2)

  addition of aspartate,

 loss of fumarate (a net gain of an amine),

 addition of another carbon (from N10N10-

formyltetrahydrofolate), and

 closing of the second ring to form inosine monophosphate

(IMP).

 IMP is a branch point for the synthesis of the adenine and guanine.

IMP can then be transformed either to GMP by IMP dehydrogenase ,

or to AMP by adenylosuccinate synthetase .

Purine Salvage Pathway

A salvage pathway is a pathway in which nucleotides are synthesized

from intermediates in the degradative pathway for nucleotides.

Overview of the Pathway

 Bases from degraded nucleic acids can be converted back into purine

nucleotides via the salvage pathways.

 Hypoxanthine can be combined with PRPP (which acts as the donor of

ribose-5 phosphate) to form IMP in a reaction catalyzed by

Hypoxanthine-guanine phosphoribosyltransferase (HGPRT).

 IMP can subsequently be transformed into AMP or GMP via the last

few steps of the pathway of de novo purine synthesis.

 HGPRT also catalyzes the reaction which combines PRPP with

guanine to form GMP.

 Adenine phosphoribosyltransferase converts adenine and PRPP to

form AMP.
 Pyrimidine biosynthesis

 Pyrimidine is a crucial part of DNA, RNA, and many other

significant biological components.

 The basic process of pyrimidine synthesis supplies the essential

building blocks for the synthesis of DNA and RNA.

 Pyrimidine nucleotide synthesis needs to be balanced for cells to

grow, proliferate, and perform normally as a whole.

 Pyrimidine synthesis disruptions can have serious effects on

cellular homeostasis and play a role in the emergence of a number

of illnesses.

 In most species, including humans, the de novo pyrimidine

synthesis process occurs in the cytoplasm of cells. Within the

cytoplasm of prokaryotes like bacteria, the complete procedure

takes place.

 The first three enzymatic stages of the process take place in the

cytoplasm of eukaryotes, which includes animals and plants,

whereas the remaining reactions happen in the mitochondria.

Reactions Involved in De novo pyrimidine synthesis

Simple precursors are transformed into the pyrimidine nucleotides,

cytosine, and uracil, by a sequence of enzyme events in the de

novo pyrimidine synthesis pathway. There are six main stages in

the route.

1. Carbamoyl Phosphate Synthesis

The pathway commences with the conversion of bicarbonate and

glutamine into carbamoyl phosphate.

This reaction is catalyzed by the enzyme carbamoyl phosphate

synthetase-II (CPS-II), which requires ATP as an energy source.

Reaction: Bicarbonate + Glutamine + 2 ATP → Carbamoyl

phosphate + Glutamate + 2 ADP + Pi

2. Formation of Carbamoyl Aspartate

In this step, carbamoyl phosphate reacts with aspartate to produce

carbamoyl aspartate. The enzyme aspartate transcarbamoylase

(ATCase) catalyzes this reaction.

Reaction: Carbamoyl phosphate + Aspartate → Carbamoyl

aspartate + Pi
3. Formation of Dihydroorotate

Carbamoyl aspartate is then converted into dihydroorotate by the

enzyme dihydroorotase.

This step does not involve the incorporation of any atoms from the

other substrates.

Reaction: Carbamoyl aspartate → Dihydroorotate + H2O

4. Formation of Orotate

Dihydroorotate is oxidized to orotate by the enzyme dihydroorotate

dehydrogenase. This reaction requires the presence of a flavin

mononucleotide (FMN) cofactor.

Reaction: Dihydroorotate + NAD+ → Orotate + NADH + H+

5. Formation of Orotidine 5′-Monophosphate (OMP)

Orotate is converted into orotidine 5′-monophosphate (OMP)

through decarboxylation and the subsequent addition of a

phosphate group.

The enzyme orotate phosphoribosyltransferase (OPRT) catalyzes

this reaction.

Reaction: Orotate + 5-Phosphoribosyl-1-pyrophosphate (PRPP) →

OMP + Pyrophosphate

6. Conversion of OMP to Uridine 5′-Monophosphate (UMP)

OMP is finally converted into uridine 5′-monophosphate (UMP)

through the addition of a ribose phosphate group.

This reaction is facilitated by the enzyme orotidine 5′-phosphate

decarboxylase (OMP decarboxylase).

Reaction: OMP → Uridine 5′-monophosphate (UMP) + CO2

7. UMP is phosphorylated to UTP in two steps. CTP is formed by

CTP synthetase by transferring an amino group from glutamine to

UTP. Pyrimidine biosynthesis is regulated by feedback inhibition

of the first enzyme carbamoyl phosphate synthetase by the end

products UMP, UDP, and UTP of the pathway.

For thymidine biosynthesis, Uridine may also be a source of

thymidine (which is a nucleoside of thymine). In order to

synthesize thymidine, uridine is reduced first to deoxyuridine (by

the enzyme ribonucleotide reductase). After which, it is methylated

by the enzyme thymidylate synthase to form thymidine.

Synthesis of CTP The transfer of an amino group from glutamine

to UTP by CTP synthetase leads to the synthesis of CTP.

Pyrimidine Synthesis via Salvage Pathways

Cells contain salvage routes for the production of pyrimidine

nucleotides in addition to de novo synthesis.

Salvage processes reduce the need for de novo synthesis by

recycling nucleobases produced during DNA and RNA

breakdown.

Through the use of certain salvage enzymes, the salvage route

involves the recovery of free pyrimidine bases such as cytosine,

uracil, and thymine.

The actions of cytidine deaminase and uridine phosphorylase

enzymes, respectively, can salvage cytidine and uridine.

By converting cytidine and uridine into uracil, these enzymes

enable the synthesis of pyrimidine nucleotides from scratch.

The enzyme thymidine kinase, which phosphorylates thymidine to

create thymidine monophosphate (TMP), is used in a separate

mechanism to salvage thymidine.