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Nucleotide Structure
Numbering
Carbon and nitrogen atoms in the rings of the base and the sugar are numbered separately
Atoms in the rings of the bases are numbered
o 1-6 in PYRIMIDINE and 1-9 in PURINE
Carbons in the pentose are numbered 1-5
Biomedical Importance
Biosyn regulated and coord’d by feedback mechanisms
Production varies, maybe in accord w/physio demand
Crazy image of nucleotide pathways: see manual, pg 101
Fate of Ingested
Nucleic acids and nucleotides
NOT incorporated directly into tissue nucleic acids
Degraded in GIT
o First into MONOnucleoTides by ribonucleases, deoxyribonucleases and polycleotidases
o Second into nucleoSides by nucleotidases and phosphatases via hydrolysis
o Then, absorbed
or
o Further degradaded into purine and pyrimidine bases by intestinal phosphorylase
Purine bases –oxidized uric acid
o Can be absorbed and excreted in urine
Degradation of dietary nucleic acids in the small intestine (see image pg 116)
Salvage Pathways
Salvage pathways are important for cells that don’t have PRPP glutamyl amido ransferase – i.e. RBCs
Ribose-1-PO4
PRPP
Pi
PPi Nucleoside
NMP
Kinase
ATP
NDP
NMP
Kinase
ATP
NMP
NMP NTP
Step 2: 5-Phosphoribosyl-b-1-amine synthesis from a-PRPP, glutamine, and H2O by glutamine phosphoribosyl
pyrophosphate amidotransferase.
The amide of glutamine replaces the pyrophosphate group attached to carbon 1 of PRPP.
Glutamine phosphoribosyl pyrophosphate amidotransferase
o Inhibited by the
purine 5’-
nucleotides
AMP,GMP and
IMP, the end
products of this
pathway.
o This is the
committed step
in purine
nucleotide
biosynthesis
Enzymes involved
1. 6-Mercaptopurine (6MP)
Antitumor drug
Analogue of adenine
Metabolized to a ribonucleotide by the APRT salvage pathway inhibiting the conversion of IMP to GMP or AMP
Also inhibits the rate limiting step of the purine de novo pathway
Simultaneous administration with allopurinol potentiates its effect as 6-Mercaptopurine will not be degraded and
therefore will delay its inactivation.
3. Azaserine
Analogue of glutamine
Inhibits
o incorporation of N3 and N9 into the purine ring (de novo biosynthesis
o formation of GMP from IMP, CTP from UTP-all reactions requiring the entry of glutamine.
Hyperuricemia charac’d by frequent episodes of uric acid lithiasis & a bizarre syndrome of self mutilation
Complete deficiency of hypoxanthine-guanine phosphoribosyl transferase (HGPRT)
o Inability to salvage hypoxanthine or guanine
levels of PRPP purine overproduction Excessive production of uric acid and IMP and GMP and de
novo syn
Mutations include deletions, frameshift mutations, base substitutions, and aberrant mRNA splicing.
Hypouricemia
Hypouricaemia and excretion of hypoxanthine and xanthine are associated with xanthine oxidase deficiency
Lecture, Manual 8 mra
Med1C Villiran – Nucleotide Metabolism Biochem 1b
Amination of UTP
1. By CTP synthase (synthetase, saw both used)
2. Nitrogen provided by glutamine
Orotic acidurias
Charac’d by high levels of urinary orotic acid, retarded growth and megaloblastic anemia
Treat w/uridine and high pyrimidine diet
One type that accompanies Reye’s syndrome
o Probably consequence of the inability of severely damaged mitochondria to utilize carbamoyl phosphate
o w/c then becomes available for cytosolic overproduction of orotic acid
Prevents formation OMG UMP UTP w/c CTP UMP UTP w/c CTP
of
Inhibiting Step 2 enzyme of Purine de novo – ATCase, per CTP syn
manual tho I don’t understand why
Excess carbamoyl phosphate exits to the cytosol, where it stimulates pyrimidine nucleotide biosyn
The resulting mild orotic aciduria is increased by high nitrogen containing food.
Manifest: Protein intolerance and hyperammonemia w/hepatic encephalopathy
Table (from Manual) of Pyrimidine Disorders
Clinical Disorder Defective Enzyme Characteristics Inheritance Pattern
Beta-Aminoisobutyric Transaminase No symptoms; frequent in Autosomal recessive
aciduria Asians
Orotic aciduria, Type I Orotate Orotic acid crystalluria
phosphoribosyltransferase Failure to thrive
and orotidylate Megaloblastic anemia
decarboxylase Immunodef
Tx:Remission w/oral uridine
Orotic aciduria, Type II Orotidylate decarboxylase Orotidunria
Orotic aciduria
Megaloblastic anemia
Tx:Remission w/oral uridine
Ornithine transcarbamoylase Ornithine transcarbamoylase Protein intolerance X-linked recessive
deficiency (OTCD) Hepatic encephalopathy
Mild orotic aciduria
Allopurinol
o alternative substrate for orotate phosphoribosyltransferase
o competes with orotic acid
Resulting nucleotide product also inhibits orotidylate decarboxylase orotic aciduria and ortidinuria
6-Azauridine
o following conversion to 6-azauridylate, also competitively inhibits orotidylate decarboxylase
o enhancing exretion of orotic acid and orotidine
2. 5 Flurouracil (5 FU)
An analogue of thymine
Treatment of solid tumors
o It is converted to the monophosphate nucleotide form via the salvage pathway.
o Eventually, converted to the deoxynucleotide form and binds to thymidylate synthetase inhibiting the
formation of TMP.
o As a deoxytriphosphate form, it can be incorporated into RNA and inhibits the formation of mature RNA (a
step important in translation)
3. 5-Iodouracil
Analogue of thymidine
When incorporated into DNA bonds with C rather than A, thus causing misreading of the strand.
Ribonucleotides
Use as building blocks in RNA synthesis & nucleotide carriers of other compound
Nucleotides required for DNA syn are 2’ – deoxyribonucleotides,
o w/c are produced from ribonucleoside diphosphate
Ribonucleotide reductase
Responsible for maintaining a balance supply of the deoxyribonuclotides required for DNA synthesis.
Note: in add’n to the single active site, there are two allosteric sites on for regulating activity
prevent folic acid syn in bacterial cell (but not man) inhibiting TMP syn and DNA rep in bacterial
cells