An In Vivo Yeast-Based Activity Assay for the PPIP5K Family.
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.
Alcázar-Román AR, Fleig U