Rosenbach, Hannah and Walla, Eva and Cutsail, George E and Birrell, James A and Pascual-Ortiz, Marina and DeBeer, Serena and Fleig, Ursula and Span, Ingrid (2021) The Asp1 pyrophosphatase from S. pombe hosts a [2Fe-2S]²⁺ cluster in vivo. Journal of Biological Inorganic Chemistry, 26 (1). pp. 93-108. DOI https://doi.org/10.1007/s00775-020-01840-w
Rosenbach, Hannah and Walla, Eva and Cutsail, George E and Birrell, James A and Pascual-Ortiz, Marina and DeBeer, Serena and Fleig, Ursula and Span, Ingrid (2021) The Asp1 pyrophosphatase from S. pombe hosts a [2Fe-2S]²⁺ cluster in vivo. Journal of Biological Inorganic Chemistry, 26 (1). pp. 93-108. DOI https://doi.org/10.1007/s00775-020-01840-w
Rosenbach, Hannah and Walla, Eva and Cutsail, George E and Birrell, James A and Pascual-Ortiz, Marina and DeBeer, Serena and Fleig, Ursula and Span, Ingrid (2021) The Asp1 pyrophosphatase from S. pombe hosts a [2Fe-2S]²⁺ cluster in vivo. Journal of Biological Inorganic Chemistry, 26 (1). pp. 93-108. DOI https://doi.org/10.1007/s00775-020-01840-w
Abstract
The Schizosaccharomyces pombe Asp1 protein is a bifunctional kinase/pyrophosphatase that belongs to the highly conserved eukaryotic diphosphoinositol pentakisphosphate kinase PPIP5K/Vip1 family. The N-terminal Asp1 kinase domain generates specific high-energy inositol pyrophosphate (IPP) molecules, which are hydrolyzed by the C-terminal Asp1 pyrophosphatase domain (Asp1<sup>365-920</sup>). Thus, Asp1 activities regulate the intracellular level of a specific class of IPP molecules, which control a wide number of biological processes ranging from cell morphogenesis to chromosome transmission. Recently, it was shown that chemical reconstitution of Asp1<sup>371-920</sup> leads to the formation of a [2Fe-2S] cluster; however, the biological relevance of the cofactor remained under debate. In this study, we provide evidence for the presence of the Fe-S cluster in Asp1<sup>365-920</sup> inside the cell. However, we show that the Fe-S cluster does not influence Asp1 pyrophosphatase activity in vitro or in vivo. Characterization of the as-isolated protein by electronic absorption spectroscopy, mass spectrometry, and X-ray absorption spectroscopy is consistent with the presence of a [2Fe-2S]<sup>2+</sup> cluster in the enzyme. Furthermore, we have identified the cysteine ligands of the cluster. Overall, our work reveals that Asp1 contains an Fe-S cluster in vivo that is not involved in its pyrophosphatase activity.
Item Type: | Article |
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Uncontrolled Keywords: | Biocatalysis; Cysteine; Cytoskeletal Proteins; Iron-Sulfur Proteins; Multifunctional Enzymes; Mutation; Phosphotransferases (Alcohol Group Acceptor); Pyrophosphatases; Schizosaccharomyces; Schizosaccharomyces pombe Proteins |
Divisions: | Faculty of Science and Health Faculty of Science and Health > Life Sciences, School of |
SWORD Depositor: | Unnamed user with email elements@essex.ac.uk |
Depositing User: | Unnamed user with email elements@essex.ac.uk |
Date Deposited: | 23 Nov 2022 11:51 |
Last Modified: | 07 Aug 2024 20:16 |
URI: | http://repository.essex.ac.uk/id/eprint/34059 |
Available files
Filename: The Asp1 pyrophosphatase from S. pombe hosts a [2Fe-2S]sup2+sup cluster in vivo.pdf
Licence: Creative Commons: Attribution 3.0