Nucleotide Metabolism Flashcards
What is a nucleoside?
base linked to 1’ of ribose
What is a nucleotide?
nucleoside esterified to a phosphate at 5’ of ribose ring
What two components are fused to make a pyrimidine?
purine and imidazole ring
Where are the enzymes for purine synthesis located?
cytoplasm
At what base is purine synthesis initiated?
PRPP
5-phosphoribosyl-1-pyrophosphate
What is the first step in purine synthesis? What enzyme catalyzes this reaction?
generation of PRPP
PRPP Synthetase
Is PRPP synthesis specific to purine synthesis?
No.
PRPP is required for both purine and pyrimidine synthesis
What is PRPP derived from?
ribose-5-phosphate
What is the committed step of de novo purine synthesis? What enzyme catalyzes this reaction?
formation of 5-phosphoribosylamine (5-PRA)
enzyme = amidophosphoribosyltransferase
What is the first purine to be produced during de novo synthesis?
inosine-5-monophosphate
IMP
IMP serves as a precursor for what two other purines?
AMP and GMP
What donate carbon and nitrogen atoms to purine synthesis?
amino acids
Specifically, what amino acids donate carbon and nitrogen to purine synthesis?
glycine, glutamine and aspartate
What does CO2 provide in purine synthesis?
carbon and oxygen
What does N10-formyl-THF provide in purine synthesis?
donor of one-carbon
What are the roles of nucleoside monophosphate kinases and nucleoside diphosphate kinases?
generation of nucleotide triphosphates
Which, NMPK orNDPK have a more specific or more general substrate specificity?
NMPK = more specific
NDPK = more broad
Why is GTP an ‘ATP Equivalent’?
the more are readily interconverted
What are the most effective inhibitors of PRPP Synthetase?
ADP and GDP
What serves to inhibit amidophosphoribosyltransferase activity?
AMP and GMP
What serves as an allosteric activator of amidophosphoribosyltransferase?
PRPP
Where does the bulk of purine nucleotide degradation occur?
the liver
How is the degrdation of GMP initiated?
5’-nucleotidases
What is the 2nd step of guanosine degradation? What two molecules are liberated?
purine nucleoside phosphorylase
guanosine –> guanine and ribose
How does AMP degradation begin?
by the action of AMP deaminase and adenosine deaminase
What does the action of AMP deaminase produce?
IMP from AMP
What enzyme converts AMP to adenosine?
5’-nucleotidases
What enzyme converts AMP to IMP?
AMP deaminase
What enzyme converts adenosine to inosine?
adenosine deaminase
What enzyme converts IMP to inosine?
5’-nucleotidases
What enzyme converts inosine to hypoxanthine?
purine nucleoside phosphorylase
What enzyme converts hypoxanthine to xanthine?
xanthine oxidase
What enzyme converts GMP to guanosine?
5’nucleosidases
What enzyme converts guanosine to guanine?
purine nucleoside phosphorylase
What enzyme converts guanine to xanthine?
guanine deaminase
What two conditions can lead to a decrease in uric acid secretion?
real insufficiency and metabolic acidosis
What two conditions can cause an increase in nucleotide turnover that can exacerbate gout?
increased nucleotide turnover
diets rich in purine
How does alcohol consumption exacerbate gout?
alcohol consumption causes ATP turnover and increase in serum lactic acid
What are gout crystals?
sodium urate
What is it called when gout precipitates in the kidney?
uric acid urolithiasis
What is allopurinol oxidized to?
oxypurinol
How does oxypurinol aid in the treatment of gout?
oxypurinol is a competitive inhibitor of xanthine oxidase
How does pyrimidine synthesis differ from that of purine synthesis?
pyrimidines are synthesized as bases then attached to ribose
What is the first enzyme of pyrimidine synthesis?
Carbamoyl Phosphate Synthetase II
What is the product of carbamoyl synthetase II?
carbamoyl phosphate
What enzyme acts on carbamoyl phosphate? What does this produce?
Aspartate Transcarbamoylase
N-carbamoyl aspartate
What enzyme works on N-carbamoyl aspartate?
Dihydroorotase
What does dihydroorotase produce?
dihydroorotate
What enzyme of pyrimidine synthesis is expressed in the mitochondria?
Dihydroorotate Dehydrogenase
What does dihydroorotate dehydrogenase work on? What does dihydroorotase produce?
dihydroorotate
orotate
What enzyme works on orotate? What is the product of this enzyme?
orotate phosphoribosyl transferase
OMP
What enzyme works on OMP? What is the product of this enzyme?
OMP decarboxylase
UMP
What enzyme catalyzes the transfer of R5P to orotate?
orotate phosphoribosyltransferase
What is OMP also known as?
orotidine 5 monophosphate
What is CAD?
carbamoyl phosphate synthetase, aspartate transcarbamoylase, dihydroorotase
What two enzymes make up UMP synthase?
OMP decarboxylase and orotate phosphoribosyl transferase
Orotic aciduria features a defect in what enzyme?
UMP synthase
What three symptoms do patients with orotic aciduria present with?
orotic acid in urine
megaloblastic anemia
occasional defect in cellular immunity
What is the treatment for orotic aciduria?
pyrimidine supplementation
What does CTP synthase use as an amino donor?
glutamine
Where is de novo synthesis of pyrimidines most regulated?
carbamoyl phosphate synthetase II
Regarding CTP synthase, what kind of effector is CTP?
negative effector
What kind of kinetics does CTP synthase display regarding UTP levels?
sigmoidal kinetics
Activity of carbamoyl phosphate synthetase II is affected by what two molecules?
inhibited by UTP
activated by PRPP
What enzyme produces deoxyribonucleotides?
ribonucleotide reductase
What is the substrate for ribonucleotide reductase?
nucleoside 5’ Di-phosphates
What is the product of ribonucleotide reductase?
2’-deoxyribonucleotide -5’-diphosphates
How are ribonucleotide reductase levels regulated?
transcription
What is a common and important pharmacological inhibitor of ribonucleotide reductase?
hydroxyurea
Mutations in adenosine deaminase are implicated in what disease?
severe combined immunodeficiency disease
Why is adenosine deaminase believed to cause SCID?
back-up of adenosine metabolites inhibits ribonucleotide reductase
How is dTMP converted to dTDP?
dTMP kinase
What is the classic inhibitor of thymidylate synthase?
5-fluorouracil
In the body, what is 5-FU converted to?
fUMP
What form of 5-fluorouracil is a substrate for ribonucleotide reductase?
fUDP
Dephosphorylation of what producs fdUMP?
fdUDP
Specifically, how does 5-FU inhibit thymidylate synthase?
FdUMP and N5N10 THF form a permanent and irreversible complex with thymidylate synthase
Degradation of pyrimidines yield what two products?
beta-alanine
beta-aminoisobutyrate
Where are nucleosides taken up?
intestinal epithelial cells
Why is adenosine interesting?
adenosine can be directly phosphorylated by adenosine kinase to produce AMP
What does Lesch-Nyhan Syndrome result from?
defective HGPRTase
How do salvage pathways decrease de novo synthesis?
By sequestering PRPP and lowering activity of amidophosphoribosyl transferase
What is the one pyrimidine that is not salvaged in humans?
cytosine
What enzyme do fungi express that make them susceptible to analogs?
cytosine deaminase
How does probenecid work?
inhibits renal uptake of uric acid
What enzyme in pyrimidine synthesis uses PRPP?
orotate phosphoribosyltransferase
What disease is UMP synthase defective in?
orotic aciduria
How is orotic aciduria treated?
uridine administration
What does CTP synthase use as a nitrogen source?
glutamine
What is CPS II inhibited by?
UTP
What is CPS II activated by?
PRPP
What is CTP synthase inhibited by?
CTP
What is CTP synthase activated by?
UTP
What is an important inhibitor of ribonucleotide reductase?
hydroxyurea
What does ribonucleotide reductase work on?
5’-nucleoside diphosphates
What enzyme is deficient in patients with SCID?
adenosine deaminase
What kind, oxy or deoxy, form of thymine exists?
Only deoxy
What is another inhibitor of thymidylate synthase?
methotrexate
How does methotrexate inhibit thymidylate synthase?
inhibitor of dihydrofolate reductase
What is beta-alanine metabolized into?
acetyl-CoA
What is dTMP metabolized into?
beta-aminoisobutyrate
What is the function of adenosine kinase?
to phosphorylate adenosine to AMP
What is the pyrimidine that IS NOT salvaged in humans?
cytosine
What do fungi express that make them vulnerable to 5-fluorocytosine?
cytosine deaminase
What does cytosine deaminase synthesize from 5-fluorouracil?
FUMP
What structures provide carbon and nitrogen for IMP synthesis?
glycine, glutamine and aspartate
What form of THF is used for purine synthesis?
N10-formyl-THF
What is the ATP consumption required to synthesize the first purine, IMP?
four ATPs consumed
What is the rate limiting step of AMP synthesis?
adenylosuccinate synthetase
What it the rate limiting step of IMP synthesis?
IMP dehydrogenase
What is amidophosphoribosyltransferase inhibited by?
AMP and GMP
What is PRPP synthetase inhibited by?
ADP and GDP
What serves as an allosteric activator of amidophosphoribosyltransferase?
PRPP
What is the function of AMP deaminase?
AMP –> IMP
What is the function of adenosine deaminase?
adenosine –> Inosine
What is more common, reduced excretion of uric acid or increased production of uric acid?
reduced excretion
How does alcohol contribute to gout?
increases serum lactic acid
uric acid competes with other organic acids for excretion
What enzyme converts dUMP to dTMP?
thymidylate synthase
What products get degraded to beta-alanine?
dCMP, CMP, UMP, uracil
What products get degraded to beta-aminoisobutyrate?
dTMP, thymine
Pyrimidine degradation requires what reducing agent?
NADPH
How are beta-alanine and beta-isobutyrate excreted?
urine
What is the function of adenosine kinase?
Adenosine –> AMP
What enzyme is deficient in Lesch-Nyhan Syndrome?
HGPRTase
What enzyme is responsible for pyrimidine salvage?
Pyrimidine Phosphoribosyltransferase
What is the one pyrimidine that is not salvaged in humans?
cytosine
How are the activities of HGPRTase and APRTase regulated?
the activity is inhibited by their products (IMP, GMP, AMP)
What is beta-alanine metabolized to?
acetyl-CoA
What is beta-aminoisobutyrate metabolized into?
succinyl-CoA
What metabolite of pyrimidine degradation is unique to pyrimdine degradation?
beta-aminoisobutyrate