[17] Nucleotide synthesis from 5-phosphoribosylpyrophosphate: Base + PRPP ⇄ nucleoside 5′-phosphate + PP
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Publisher SummaryThis chapter describes the nucleotide synthesis from 5-phosphoribosylpyrophosphate (PRPP). The synthesis of nucleotides from PRPP takes place in one step with the concomitant formation of pyrophosphate (PP). Enzymes catalyzing reactions of this type have been termed nucleotide pyrophosphorylases. The reaction is distinct from another widely distributed pathway of nucleotide synthesis which proceeds through the nucleoside, followed by phosphorylation to the nucleotide level. AMP formation is determined by measuring the incorporation of C14-adenine into a barium–ethanol insoluble fraction, obtained after the addition of carrier AMP. Residual C14-adenine is removed, and the radioactivity of the insoluble barium adenylate is determined, the latter being a measure of the AMP formed. The purified yeast enzyme is contaminated with inorganic pyrophosphatase and a pyrophosphorylase for IMP and GMP. This latter activity can be preferentially absorbed to calcium phosphate gel by increasing the amounts of gel used in the last step of the purification procedure. The assay method for uridine 5′-phosphate pyrophosphorylase is based on the conversion of C14-uracil to UMP.

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