Alcázar-Román Abel R, Fleig Ursula
Eukaryotic Microbiology, Heinrich-Heine-University, Düsseldorf, Germany.
Methods Mol Biol. 2025;2972:205-219. doi: 10.1007/978-1-0716-4799-8_15.
The highly conserved eukaryotic PPIP5K family of bifunctional enzymes regulate cellular levels of 1,5-InsP8, a high-energy molecule involved in a multitude of biological processes. Members of this family contain two opposing activities: an N-terminus ATP-grasp kinase domain synthesizing 1,5-InsP8 and a C-terminus pyrophosphatase domain degrading 1,5-InsP8. While biochemical characterization of members of this family is vital, we present a complementary, simple, and very fast in vivo assay in the fission yeast Schizosaccharomyces pombe, for the functional assessment of kinase and pyrophosphatase activities of PPIP5K family members of any species.
高度保守的真核双功能酶PPIP5K家族调节1,5-InsP8的细胞水平，1,5-InsP8是一种参与多种生物过程的高能分子。该家族成员具有两种相反的活性：N端ATP结合激酶结构域合成1,5-InsP8，C端焦磷酸酶结构域降解1,5-InsP8。虽然对该家族成员进行生化特性分析至关重要，但我们在裂殖酵母粟酒裂殖酵母中提出了一种互补、简单且非常快速的体内检测方法，用于对任何物种的PPIP5K家族成员的激酶和焦磷酸酶活性进行功能评估。
