Non-typical nucleoside analogs as fluorescent and fluorogenic indicators of purine-nucleoside phosphorylase activity in biological samples,Analytica Chimica Acta

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Non-typical nucleoside analogs as fluorescent and fluorogenic indicators of purine-nucleoside phosphorylase activity in biological samples
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2020-12-01 , DOI: 10.1016/j.aca.2020.09.018
A Stachelska-Wierzchowska ₁ , J Wierzchowski ₁

Affiliation

Several novel non-typical nucleoside analogs were examined as potential fluorescent indicators of purine-nucleoside phosphorylase (PNP) activity in human blood. The substrates included N7-riboside of 8-aza-2,6-diaminopurine, N6-riboside of 1,N6-etheno-adenine and N2-riboside of N2,3-etheno-2-aminopurine. Reaction rates and apparent Michaelis' constants were determined in 1000-fold blood lysates and compared with those for reference compounds, guanosine and 7-methylguanosine. It was concluded that the most promising for assaying human PNP in biological material was N6-riboside of 1,N6-etheno-adenine and N2-riboside of N2,3-etheno-2-aminopurine was optimal for the E. coli PNP, both offering at least 10-fold improvement in sensitivity relative to conventional assays. Other potential applications of this approach are discussed.

中文翻译：

非典型核苷类似物作为生物样品中嘌呤核苷磷酸化酶活性的荧光和荧光指示剂

几种新的非典型核苷类似物被检查为人体血液中嘌呤核苷磷酸化酶 (PNP) 活性的潜在荧光指示剂。底物包括8-aza-2,6-二氨基嘌呤的N7-核苷、1,N6-乙烯-腺嘌呤的N6-核苷和N2,3-乙烯-2-氨基嘌呤的N2-核苷。在 1000 倍的血液裂解物中测定反应速率和表观米氏常数，并与参考化合物鸟苷和 7-甲基鸟苷进行比较。得出的结论是，在生物材料中测定人 PNP 最有希望的是 1,N6-乙烯-腺嘌呤的 N6-核苷和 N2,3-乙烯-2-氨基嘌呤的 N2-核苷对于大肠杆菌 PNP 是最佳的，两者与传统检测相比，灵敏度至少提高 10 倍。讨论了这种方法的其他潜在应用。

更新日期：2020-12-01

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