Development, validation and application of a 96-well enzymatic assay based on LC-ESI-MS/MS quantification for the screening of selective inhibitors against Mycobacterium tuberculosis purine nucleoside phosphorylase.

@article{Cattaneo2016DevelopmentVA,
  title={Development, validation and application of a 96-well enzymatic assay based on LC-ESI-MS/MS quantification for the screening of selective inhibitors against Mycobacterium tuberculosis purine nucleoside phosphorylase.},
  author={Giulia Cattaneo and Daniela Ubiali and Enrica Calleri and Marco Rabuffetti and Georg C H{\"o}fner and Klaus T. Wanner and Marcela Cristina de Moraes and Leonardo K. Martinelli and Di{\'o}genes Santiago Santos and Giovanna Speranza and Gabriella Massolini},
  journal={Analytica chimica acta},
  year={2016},
  volume={943},
  pages={
          89-97
        }
}
Mycobacterium tuberculosis (Mtb) purine nucleoside phosphorylase (PNP, EC 2.4.2.1) has been identified as a target for the development of specific inhibitors with potential antimycobacterial activity. We hereby described the development and validation of a new 96-well LC-ESI-MS/MS method to assess the inhibition activity of nucleoside analogues towards MtbPNP and the human PNP (HsPNP). Enzyme activity was determined by monitoring the phosphorolysis of inosine (Ino) to hypoxanthine (Hpx). The… 
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