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Almost all cell types have the ability to synthesize purine and pyrimidine
nucleotides from low molecular weight precursors in amounts sufficient
for their own needs.
The de novo pathways are almost identical in all organisms.






Most organisms have the ability to synthesize nucleotides from
nucleosides or bases that become available through the diet or from
degredation of nucleic acids.
In animals, the extracellular hydrolysis of ingested nucleic acids
represents the major route by which bases become available.
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blue-catabolism
red-salvage pathways

endonucleases:
pancreatic RNAse
pancreatic DNAse

phosphodiesterases:
usually non-specific
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Enzyme inhibited by AMP, ADP, and GDP. In _ , expression is repressed
by PurR repressor bound to either guanine or hypoxanthine.

Roles of PRPP: his and trp biosynthesis, nucleobase salvage pathways, de


novo synthesis of nucleotides
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þ , the reaction is driven to the right by the action of pyrophosphatase

Shown: HGPRT, cells also have a APRT.


        
 
     

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The base in IMP is called
hypoxanthine
Note: purine ring built up at
nucleotide level.
precursors:
glutamine (twice)
glycine
N10-formyl-THF (twice)
HCO3
aspartate
In vertebrates, 2,3,5 catalyzed
by trifunctional enzyme,
6,7 catalyzed by bifunctional
enzyme.
 
 
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components of the complex:


GAR transformylase (3)
AICAR transformylase (9)
serine hydroxymethyl transferase, trifunctional formylmethenyl-
methylene-THF synthase (activities shown with asterisk)


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G-1: IMP dehydrogenase


G-2: XMP aminase
A-1: adenylosuccinate
synthetase
A-2: adenylosuccinate lyase

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GTP used to make AMP,
ATP used to make GMP.
Also, feedback inhibition by
AMP and GMP.


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Conversion to diphosphate involves specific kinases:

GMP + ATP <-------> GDP + ADP #



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AMP + ATP <-------> 2 ADP $


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Conversion to triphosphate by Nucleoside diphosphate kinase (NDK):

GDP + ATP <------> GTP + ADP G0¶= 0

ping pong reaction mechanism with phospho-his intermediate.

NDK also works with pyrimidine nucleotides and is driven by mass action.
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AMP deamination in muscle, hydrolysis in other tissues.


Xanthine oxidase:contains FAD, molybdenum, and non-
non-heme iron.

In primates, uric acid is the end product, which is excreted.


  




    


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The three defects shown each result in elevated de novo purine biosynthesis
 
   


 

Allopurinol is an analogue of hypoxanthine that strongly inhibits


xanthine oxidase. Xanthine and hypoxanthine, which are soluble, are
accumulated and excreted.
 
   
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  Severe HGPRT deficiency
( )|
 
( )
In addition to symptoms of gout, patients display severe behavioral
disorders, learning disorder, aggressiveness and hostility, including self-
directed. Patients must be restrained to prevent self-mutilation. Reason
for the behavioral disorder is unknown.
X-linked trait (HGPRT gene is on X chromosome).

&"' lack of adenosine


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deaminase (ADA).
Lack of ADA causes accumulation of deoxyadenosine. Immune cells,
which have potent salvage pathways, accumulate dATP, which blocks
production of other dNTPs by its action on ribonucleotide reductase.
Immune cells can¶t replicate their DNA, and thus can¶t mount an
immune response.
    

Pyrimidine ring is assembled as the free base, orotic acid, which is


them converted to the nucleotide orotidine monophosphate (OMP).

The pathway is unbranched. UTP is a substrate for formation of


CTP.
    

1: carbamyl phosphate
synthase
2: aspartate
transcarbamylase
3: dihydroorotase
4: dihydroorotate DH
5: orotate
phosphoribosyl
tranferase
6: orotidylate
decarboxylase
7: UMP kinase
8: NDK
9: CTP synthetase

CAD=1,2,3
5 +6=single protein
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One enzyme, ribonucleotide reductase,


reduces all four ribonucleotides to their
deoxyribo derivitives.
A free radical mechanism is involved
in the ribonucleotide reductase
reaction.

There are three classes of ribonucleotide


reductase enzymes in nature:
Class I: tyrosine radical, uses NDP
Class II: adenosylcobalamin. uses NTPs
(cyanobacteria, some bacteria,
Euglena).
Class III: SAM and Fe-S to generate
radical, uses NTPs.
(anaerobes and fac. anaerobes).
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