Purine Metabolism

Raymond B. Birge, PhD

Biochemistry & Molecular Biology
Nucleotides play key roles in many, many cellular processes

1. Activated precursors of RNA and DNA

2. Adenine nucleotides are components of three major

co-enzymes, NAD, FAD, and CoA

3. Nucleotide derivatives are activated intermediates in

biosynthetic processes (UDP-glucose, SAM)

4. Serve as metabolic regulators (e.g cAMP and

the activation of cell signaling).

5. Serve as major currency of energy in all cells

(ATP and GTP).

6. Many metabolic diseases have their etiology in nucleotide

metabolism.
 The nomenclature of purines depends
on their linkage to a pentose

Adenine Adenosine Adenosine Monophosphate

Base Nucleoside* Nucleotide
Base Base (P04 ester)

* when the base is purine, then the nucleoside ends in OSINE (AdenOSINE, GuanOSINE, InOSINE)
The active forms of nucleotides in biosynthesis
and energy conversions are di-and tri-phosphates

Nucleoside Monophosphate Kinase

(i) CMP + ATP CDP + ADP

Nucleoside Diphosphate Kinase

(ii)
XDP + YTP XTP + YDP
 What do nucleosides and nucleotides do?

Purine binding proteins (“the purine proteome”) comprise a family of 3-4,000

Proteins and as much as 50% of all druggable targets in biology.

Kinases
Helicases
Reductases
Transferases
Synthetases
Dehydrogenases
Chaperones
Metabolic Enzymes
DNA and RNA processing
Etc
 The nomenclature of purines depends
on their linkage to a pentose

Adenine Adenosine Adenosine Monophosphate

Base Nucleoside* Nucleotide
Base Base (P04 ester)

* when the base is purine, then the nucleoside ends in OSINE (AdenOSINE, GuanOSINE, InOSINE)
 Nucleoside Function in extracellular signal transduction

Adenosine nucleoside-increased
during ATP degradation.

Released in cells when there

is low O2 concentration

Binds to purinogenic receptors

A1, A2A, A2B, A3

Slows the heart down, at the same

time increases capillary dilation

Caffeine is a adenine derivative, and

antagonizes the effects of adenine.
Cyclic nucleotides are important mediators for
Intracellular signal transduction

10-10 M

ATP cAMP

PKA
P P
phosphorylase glycogen
kinase synthase

P
phosphorylase
Modified nucleotide mono and di phosphates have important in electron transfer and
Redox control
RIBONUCLEOTIDE REDUCTASE

1. Complex enzymatic reaction whereby electrons

are transferred from NADPH through a series of
sufhydryl groups at the catalytic site of Ribonucleotide
Reductase.

2. Active site of RR contains thioredoxin, a 12 kD

proteinwith two exposed cysteines,
which become oxidized.

3. This ultimately allows for the reduction of ribose.

REGULATION

1. Based on the response to cellular need for dATPs.

dATP is general inhibitor
ATP is a general activator
Nucleotides are linked by 5’ to 3’ phosphodiester bonds
to generate DNA and RNA
 Structures of Common Purine Bases.

H= 6 oxy purine A= 6 amino purine

X= 2,6 dioxy purine G= 2 amino, 6-oxy purine
 Hypoxanthine is an intermediate for Adenine and Guanine

(N source)
Aspartate

(N source)
Glutamine

The common mechanistic them for the conversion of A and G is the conversion of a
carbonyl oxygen to an amino group
There are two basic mechanisms to
generate purines and pyrimidines

1. DE NOVO BIOSYNTHETIC PATHWAYS

(building the bases from simple building blocks)

2. SALVAGE PATHWAYS
(the reutilization of bases from dietary
or catabolic sources)
 The biosynthesis of purine (A and G) begins with the
synthesis of the ribose-phosphate

Pentose phosphate
pathway

Ribose phosphate pyrophospho-KINASE

Oxidative Stages of Pentose Phosphate Pathway
Glucose
ATP
Hexokinase
PPi

Glucose 6 phosphate
NADP G6P-DH
NADPH

6-Phosphogluconolactone
H20

6-Phosphogluconate
NADP
6PG-DH
C02, NADPH

Ribulose-5-phosphate
The major regulatory step in purine biosynthesis is the conversion
of PRPP to 5-Phosphoribosyl-1-amine

*
Glutamine Glutamate

PRPP PPi
Amidophosphoribosyl
transferase

Amidophosphoribosyl transferase is an important regulatory enzyme in

purine biosynthesis. It is strongly inhibited by the end products IMP, AMP, and GMP.
This type of inhibition is called FEEDBACK INHIBITION.
Several amino acids are utilized in purine biosynthesis,

IMP is the precursor for both AMP and GMP,

the base is also called hypoxanthine
 Conversion of Hypoxanthine to Adenine/Guanine.

(N source)
Aspartate

(N source)
Glutamine

The common mechanistic theme for the conversion of A and G is the

conversion of a carbonyl oxygen to an amino group
 Purines:where do the atoms come from?

Purine intermediates include:

1. Glycine
2. 1 C units of 5,10 mTHF
3. Glutamine
4. Asparate
 The regulation of purine biosynthesis is a
classic example of negative feedback
Inhibited by AMP

AMP
Ribose Phosphoribosyl
PRPP IMP
5-phosphate amine
GMP

Inhibited by IMP,
AMP, and GMP
Inhibited by GMP
Stages of nucleotide metabolism

Endonuclease Nucleic Acid

Synthesis

Nucleoside
triphosphate
Phosphodiesterase

Nucleoside monophosphates
(Mononucleosides)
 Endonuclease Nucleic Acid
Synthesis

Nucleoside
triphosphate
Phosphodiesterase

H20 Nucleoside monophosphates

Nucleotidases
(Mononucleosides)
PPi
Pi
ADP
Phosphoribosyl
Nucleosides Nucleoside kinase
ATP
transferases
Pi

Phosphorylases PRPP
Ribose-1-P Nucleobases

Uric Acid (purines)

 Nucleotidase

Phosphorylase

Cytosine Cytidine Cytidine Monophosphate

Base Nucleoside* Nucleotide

Base Base (P04 ester)
 Gout-Gouty Arthritis
(a metabolic condition of abnormal purine metabolism)
 Salvage pathways for the re-utilization of purines;

There are 2 salvage enzymes with different specificity;

1. Adenine phosphoribosyl transferase

2. Hypoxanthine-guanine phosphoribosyl transferase
O O
O P O CH2 O P O CH2 A
O PPi Base O
OH + (ie Adenine) OH + PPi
OH OH OH OH

A-PRT

PRPP + Adenine Adenylate

HG-PRT
PRPP + Guanine Guanylate
 What happens in gout?
Inhibited by AMP

AMP

Ribose
5-phosphate
PRPP Phosphoribosyl
amine
IMP

GMP

Inhibited by IMP,
AMP, and GMP
Inhibited by GMP

1. Negative regulation of PRPP Synthatase & PRPP Amidotransferase is lost

2. PRPP levels are increased because of defects in salvage pathways

Therefore, there is net increase in biosynthetic/degradation pathways!!

Purines in humans are degraded to Urate

Important points:
1. Nucleotides are constantly
undergoing turnover!

2. There are many enzymes involved;

Nucleotidases
Nucleoside phosphorylases
Deaminases
Xanthine oxidases

3. the final common intermediate in

humans is Urate, which is excreted.

4. there are several metabolic disorders

resulting from defects in purine
catabolism.
Catabolism of Adenosine and Guanosine to Uric acid
 GOUT (Gouty Arthritis): A defect of purine metabolism

Serum Uric Acid Levels Incidence of Gout

(mg/dl) (% of cases)

>9.0 ~10%
7-9 0.5-3.5%
<7.0 0.1%

Guanine Hypoxanthine Allopurinol:

a. decrease urate
xanthine oxidase b. increase xanthine &
hypoxanthine
Xanthine Urate c. decrease PRPP
xanthine oxidase
 SCID-Severe Combined Immunodeficiency Syndrome

Autosomal recessive disorder

AMP Mutations in ADA
H20
Nucleotidase Infants subject to bacterial,
candidiasis, viral, protazoal
Pi infections
Adenosine Both T and B cells are significantly
H20 reduced (dATP is toxic)
Adenine deaminase*
NH3 1995-AdV expressing ADA was
successfully employed as gene
Inosine Hypoxanthine therapy strategy
Disorders of Purine Metabolism:

Disorder Defect Comments

Gout PRPP synthase/ Hyperuricemia
HGPRT

Lesch Nyhan lack of HGPRT Hyperuricemia

syndrome

SCID ADA High levels of dAMP

von Gierke’s disease glucose -6-PTPase Hyperuricemia