The Asp1 pyrophosphatase from S. pombe hosts a [2Fe-2S]2+ cluster in vivo

Abstract The Schizosaccharomyces pombe Asp1 protein is a bifunctional kinase/pyrophosphatase that belongs to the highly conserved eukaryotic diphosphoinositol pentakisphosphate kinase PPIP5K/Vip1 family. N-terminal domain generates specific high-energy inositol pyrophosphate (IPP) molecules, which are hydrolyzed by C-terminal pyrophosphatase (Asp1 365−920 ). Thus, activities regulate intracellular level of class IPP control wide number biological processes ranging from cell morphogenesis chromosome transmission. Recently, it was shown chemical reconstitution 371−920 leads formation [2Fe-2S] cluster; however, relevance cofactor remained under debate. In this study, we provide evidence for presence Fe–S cluster in inside cell. However, show does not influence activity vitro or vivo. Characterization as-isolated electronic absorption spectroscopy, mass spectrometry, and X-ray spectroscopy consistent with 2+ enzyme. Furthermore, have identified cysteine ligands cluster. Overall, our work reveals contains an vivo involved its activity.