• NUCLEOTIDE METABOLISM
Nucleotide Metabolism
•Chemistry of Nucleotides
•Classfication of Nucleotides
•Function of Nucleotides
•Synthesis,degradation and Associated
diseases of purine Nucleotides
•Synthesis,degradation and Associated
diseases of pyrimidine Nucleotides
Nucleotides
Nucleotides are composed of a nitrogenous base, a pentose
mono saccharides, and one,two or three phosphate groups.
A. Nitrogenous bases: These are either purine or pyrimidines
Purines:
Adenine 6 amino purine
Guanine 2amino -6- oxy purine
Pyrimidenes:
Cytosine 2-oxy-4 amino pyrimidine
Uracil : 2.4 Dioxy pyrimidine
Thymine 5-methyl pyrimidine
B. Sugar: Is either Ribose or Deoxy Ribose
Sugar + Nitrogenous base = Nuleoside
Therefore Nucleosides can be either
ribonucleoside or Deoxyribonucleoside e.g
• Adenine + Ribose Adenosine
• Adenine + Deoxy Ribose Deoxy Adenosine
• Guanine + Ribose Guanosine
• Guanine + Deoxy Ribose Deoxy Guanosine
• Cytosine + Ribose cytidine
• Cytosine + Deoxy Ribose Deoxy Cytidine
• Uracil + Ribose Uridine
• Thymine + Deoxy Ribose Deoxy Thymidine
• PHOSPHORIC ACID:
• Nucleoside + Phosphoric acid Nucleotide
• Adenosine + H3PO4 AMP
• Gaunosine + H3PO4 GMP
• Cytidine + H3PO4 CMP
• Uridine + H3PO4 UMP
• In case of Mononucleotides, a phosphoric acid
molecule forms an ester linkage with one of the OH
groups of sugar of a nucleoside.
Phosphoric acid
• In case of Ribose,
there are three such
places ( C no. 2,3,5 )
where this ester
linkage can be formed,
but in Deoxy ribose
,only two such places
i.e.3 & 5 ,because C
no.2 of sugar lack an
O2 atom.
RIBOSE & DEOXYRIBOSE
• Functions of Nucleotide
• Formation and Degradation of cyclic AMP
• Function of cyclic AMP
• Function of cyclic GMP
Folic Acids
• Active coenzyme form is called tetra hydro folate ( H4
folate)
• Tetra hydrofolate acts as a carrier in transfer of one
carbon group
• The one carbon group combines at N5 ,N10 or balanced
between nitrogen No 5 and Nitrogen No10 of folic acid
• The one carbon unit may be
1. Formyl group
2. Formamine group
3. Methyl group
4. Methenyl group
PURINE
SYNTHESIS
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)
DENOVO –SYNTHESIS
OF
PURINE NUCLEOTIDES
(i.e SYNTHESIS FROM
AMPHIBOLIC
INTERMEDIATES or synthesis
from different small compounds )
• Purines are built upon a pre-existing ribose -
5-phosphate
• Liver is the major site of synthesis
• R.B.C ,W.B.C and brain cannot synthesized
purine by Denovo synthesis.
Purines: where do the atoms come from?
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
• Antifolate Drugs and Glutamine Analogs
Block Purine Nucleotide Biosynthesis
• Six ATP utilized in de novo synthesis so
energetically expensive process.
• Amino Acid required are Glycine, Aspartic Acid
and glutamine.
• Co enzymes and Co factors are, formylated FH4,
Mg++.
• Purines & Pyrimidines are dietarily nonessential
• Purines are synth mainly in the cytosole of liver
cell.
• Nucleotides do not enter cell directly but are
converted to nucleosides by the cell
membrane nucleosidases
• In the cell nucleoside is either converted to
the Nucleotide again by a kinase enzyme or
degraded to corresponding base by the
enzyme nucleoside Phosphorylase .
SALVAGE PATHWAY FOR
PURINES
• Purines that result from the 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.
Significances of salvage pathway
• 1. Denovo synth is expensive in terms of
use of high energy phosphate bonds.
• 2. Enzyme PRPP Glutamyl amido
Transferase is not present in certain tissues
e.g Neutrophil , R.B.C and Brain cells.
• So denosynthesis cannot take place in these
tissues.
• Two pathways are available
1. One step synthesis
2. Two step synthesis
ONE STEP SYNTHESIS
Two Enzymes are involved
• i) APRT
• ii ) HGPRT
• Both enzymes utilize PRPP as a source of
the R-5-Phosphate group.
TWO STEP PATHWAY
(Nucleoside phosphorylase-
Nucleosside kinase pathway)
• Neither guanosine nor inosine kinase have
been detected
• So adenine is the only purine that may be
salvaged by the two steps pathway.
Regulation of purine synthesis
• PRPP synthetase :
Allosterically inhibited by PRPP and
AMP,GMP ,ADP,GDP,NAD ,FAD.
• Glutamin PRPP amidotransferase :
Feedback inhibition by AMP and GMP
• A proper balance between the adenine and
guanine conc maintain by adenylosuccinate
synthetase and IMP dehydrogenase
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
Ribonucleotide reductase provides the
hydrogen atoms needed for reduction from its
sulfhdryl groups.
DEGRADATION OF PURINE
NUCLEOTIDES
(Formation of uric acid)
• Xanthine oxidase contain FAD ,molybdenum
and iron and is exclusively found in liver and
small intestine.
• Molecular oxygen is reduced at each stage to
super oxide (O -2 ) which is converted to
H 2O2 by super oxide dismutase.
• Catalase cleves H 2O2 to H 2O and O2
• Allopurinol is competitive inhibitor of
Xanthine oxidase.
• Allopurinol is a best example of suiccidal
inhibition.
• Allopurinol xanthine oxidase Alloxanthine
which is a stronger inhibitor of xanthine
oxidase.
DISORDERS OF PURINE
METABOLISM
Gout
GOUT
• Gout is not a single disease.
• The term is used to describe a number of
disorders in which crystals of Mono sodium
urate monohydrate (uric acid) derived from
Hyperuricemic body fluid give rise to
• Inflammatory arthritis
• Urolithiasis
• Renal disease
3.4-------7mg/dl in males
2.4-------5.8mg /dl in Females
• Uric acid is formed by oxidation of purine
bases , which may be exogenous or
endogenous in origin.
• It is formed in the liver and excreted largely
through kidneys (2/3),some is excreted in
bile,some is converted to urea and amonia
by the intestinal bacteria.
• U.A is completely filtered at Glomerulus,
reabsorbed at prox conv Tubules and
secreted further along the Tubules.
Hyperuricemia occurs as a result
of
• A. Over production of U.A (Metabolic)
• B. Under excretion of U.A (Renal)
A.Over production
1.Genetic disorders
PRPP synthetase :
Variant forms of PRPP synthetase ,may be
i. Superactive
ii. Resistant to feed back inhibition
iii. Low Km for ribose-5-Phosphate
PRPP glutamylamidotransferase:
The lack of feedback control
HGPRT deficiency :
Von –Gierke`s disease: (G6-Phosphatase deficiency)
Fructose intolerance
2. High purine and Fructose intake:
3. Increase tissue breakdown (Treatment of
malagnancies)
4. Alcohol consumption (lactic acidosis)
B. Under Excretion
Renal diseases:
Hypertension : (Essential)
Enhances prox Tub reabsorbtion and
Depresses renal Tub secr of U.A.
Diuretic Therapy: (Similar Mechanism)
Diabetes Mellitus:( Increase Insulin. similar
mechanism)
Clinical feature of Gout
1.Metatarso Phalangeal Joint of a great Toe (70%) (Podagra).
• Other Joints may be affected .
• The effected joint is Hot,Red and swollen with shiny
overlying skin and dilated veins.
• It is severely painful and Tender.
2. Deposition of urate crystals in soft tissues (Tophaceous
gout).
3. Renal Complication
Such as pain in the lumber region due to stone formation and
the stones may cause renal failure.
Von Gierke,s Diseas
• Hyper uricemia occurs due to glucose-6-
phosphatase-deficiency
• Glucose
•
• G-6-P
• So more G-6-P will be shunted into HMP
Shunt, Resulting in more formation of
R-5-P, PRPP and Purine over production .
Lactic Acidosis
• Increased lactic acid competes with uric
acid for excretion, resulting in retention of
uric acid .
Fructose Intolerance and Gout
Fructose Fructo Kinase Fructose-I-P
Aldolase-B(Absent)
ATP ADP
Hypoxanthine D.H.
Acetone Phosphate
Xanthine
Uric Acid D-Glyceraldehyde
Gout
Disorders of Purine Catabolism
• 1. Gout
• 2.Lesch-Nyhan Syndrome
• 3.Von Gierke Disease
• 4. Hypouricemia
• 5.Adenosine Deaminase Deficiency
• 6. Purine Nucleoside Phosphorlylase
Deficiency.
SCID-Severe Combined Immunodeficiency Syndrome
AMP
Autosomal
HNucleotidase recessive
20
Pi disorder
Adenosine
Mutations in
H2Adenine
0 deaminase*
ADA
InosineNH3 Hypoxanthine
Infants
subject to
• PYRIMIDINE SYNTHESIS
Ribonucleotide reductase provides the
hydrogen atoms needed for reduction from its
sulfhdryl groups.
1. The first three enzymes of the cycle are
domains of single polypeptide chain.
2. Orotate phosphoribosyl Transferase and
OMP decarboxylase are domains of single
Polypeptide chain.
Dihydro orotate
Dehydrogenase
• Is the only mitochondiral enzyme of the
cycle .
• Contain FMN, FAD, Fe, S as Prosthetic
group and require NAD as Coenzyme
Orotic Aciduria
Mainly of two types
Type-1. Orotate phosphoribosyl transferase and OMP
decarboxylase(Both) are deficient.
• Results in accumulation of orotate in blood causing growth
retardation .Megaloblastic Anaemia.
Type-2: Deficiency of OMP decarboxylase
• Megaloblastic Aneamia .
Other causes of orotic Aciduria
1. Reye syndrome------
Damaged mitochondria
2.Deficiency of urea cycle enzyme i.e
Ornithine transcarbamyolase.
3.Drugs, e.g Allopurinol and 6-
Azauridine.
PYRIMIDINE BASE
SALVAGE
• The enzyme pyrimidine phosphoribosyl transferase
catalyzes the formation of pyrimidine nucleotide, using
PRPP as the donor of ribosyl moiety.
Pyrimidine Base
PRPP Pyrimidine phosphoribosyl
transferase.
Pyrimidine nucleotide+ PP
