1,N6-乙烯腺嘌呤及其他荧光核碱基类似物作为嘌呤核苷磷酸化酶的底物：光谱学和动力学研究
1,N6-ethenoadenine and other Fluorescent Nucleobase Analogues as Substrates for Purine-Nucleoside Phosphorylases: Spectroscopic and Kinetic Studies.
作者信息
Wierzchowski Jacek, Stachelska-Wierzchowska Alicja, Wielgus-Kutrowska Beata, Bzowska Agnieszka
机构信息
Department of Biophysics, Faculty of Food Sciences, University of Varmia & Masuria, Olsztyn. Poland.
Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw. Poland.
出版信息
Curr Pharm Des. 2017 Oct 11. doi: 10.2174/1381612823666171011103551.
BACKGROUND
Purine-nucleoside phosphorylase (PNP) is known as a tool for the synthesis of various nucleosides and nucleoside analogues. Mechanism, properties, molecular diversity and inhibitors of PNP, particularly these of pharmacological significance, are briefly characterized.
METHODS
UV and fluorescence spectroscopy was used for kinetic experiments, and HPLC chromatography for product analyses.
RESULTS
Applications of various forms of PNP to synthesis of selected fluorescent nucleosides, particularly ribosides of 1,N6-ethenoadenine and various 8-azapurines (triazolo[4,5-d]pyrimidines) are reviewed. Different specificity of various PNP forms is described towards nucleobase and analogue substrates as well as variable ribosylation sites observed in some reactions, with a possibility to further modify these features via the site-directed mutagenesis.
CONCLUSION
Present and future applications of the fluorescent or fluorogenic ribosides are discussed, with particular emphasis on biochemical and clinical analyses with improved sensitivity.
背景
嘌呤核苷磷酸化酶（PNP）是合成各种核苷和核苷类似物的一种工具。简要介绍了PNP的作用机制、特性、分子多样性及抑制剂，尤其是具有药理学意义的那些。
方法
采用紫外和荧光光谱进行动力学实验，用高效液相色谱进行产物分析。
结果
综述了各种形式的PNP在合成选定的荧光核苷中的应用，特别是1,N6-乙烯腺嘌呤和各种8-氮杂嘌呤（三唑并[4,5-d]嘧啶）的核糖苷。描述了各种PNP形式对核碱基和类似物底物的不同特异性，以及在某些反应中观察到的可变核糖基化位点，并且有可能通过定点诱变进一步改变这些特征。
结论
讨论了荧光或荧光核糖苷的当前和未来应用，特别强调了具有更高灵敏度的生化和临床分析。
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