Division of Cell Signalling, School of Life Sciences, University of Dundee, Dundee, UK.
Department of Pharmacy, University of Wolverhampton, Wolverhampton, UK.
Methods Mol Biol. 2020;2091:75-82. doi: 10.1007/978-1-0716-0167-9_6.
The diphosphoinositol polyphosphate phosphohydrolases are a subset of the Nudix hydrolase family of enzymes. As such, they metabolize a wide range of substrates, including diphosphoinositol polyphosphates (inositol diphosphates, inositol pyrophosphates), dinucleotide phosphates, nucleosides as well as 5-phosphoribosyl 1-pyrophosphate and inorganic polyphosphate. Here, we describe protocols to optimize these enzymes, with consideration to buffer composition and sample preparation and how to analyze the metabolism of these substrates using high-performance liquid chromatography, giving advice where pitfalls are commonly encountered.
二磷酸肌醇多磷酸磷酸水解酶是 Nudix 水解酶家族酶的一个亚类。因此，它们代谢的底物范围很广，包括二磷酸肌醇多磷酸（肌醇二磷酸、肌醇焦磷酸）、二核苷酸磷酸、核苷以及 5-磷酸核糖 1-焦磷酸和无机多磷酸盐。在这里，我们描述了优化这些酶的方案，考虑了缓冲液组成和样品制备，以及如何使用高效液相色谱法分析这些底物的代谢，并在常见陷阱处给出建议。
