Metal stopping reagents facilitate discontinuous activity assays of the de novo purine biosynthesis enzyme PurE.

@article{Sullivan2014MetalSR,
  title={Metal stopping reagents facilitate discontinuous activity assays of the de novo purine biosynthesis enzyme PurE.},
  author={Kelly L. Sullivan and Loredana C Huma and Elwood A. Mullins and Michael E. Johnson and T. Joseph Kappock},
  journal={Analytical biochemistry},
  year={2014},
  volume={452},
  pages={
          43-5
        }
}
The conversion of 5-aminoimidazole ribonucleotide (AIR) to 4-carboxy-AIR (CAIR) represents an unusual divergence in purine biosynthesis: microbes and nonmetazoan eukaryotes use class I PurEs while animals use class II PurEs. Class I PurEs are therefore a potential antimicrobial target; however, no enzyme activity assay is suitable for high throughput screening (HTS). Here we report a simple chemical quench that fixes the PurE substrate/product ratio for 24h, as assessed by the Bratton-Marshall… CONTINUE READING

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