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Updated: May 6, 2026

A Purification and In Vitro Activity Assay for a pppGpp Synthetase from Clostridium difficile
Published on: November 3, 2018
Two fluorogenic substrates for purine nucleoside phosphorylase, selective for mammalian and bacterial forms of the
Jacek Wierzchowski1, Alicja Stachelska-Wierzchowska1, Beata Wielgus-Kutrowska2
1Department of Biophysics, University of Varmia and Masuria, 10-719 Olsztyn, Poland.
Abstract:
Two nontypical nucleosides, 7-β-D-ribosyl-2,6-diamino-8-azapurine and 8-β-D-ribosyl-2,6-diamino-8-azapurine, have been found to exhibit moderately good, and selective, substrate properties toward calf and bacterial (Escherichia coli) forms of purine nucleoside phosphorylase (PNP). The former compound is effectively phosphorolysed by calf PNP and the latter by PNP from E. coli. Both compounds are fluorescent with λ(max) ∼ 425 to 430 nm, but the reaction product, 2,6-diamino-8-azapurine, emits in a different spectral region (λ(max) ∼ 363 nm) with nearly 40% yield, providing a strong fluorogenic effect at 350 to 360 nm.

