New insights into the function and molecular mechanisms of Ferredoxin-NADP+ reductase from Brucella ovis
Bacterial ferredoxin(flavodoxin)-NADP<sup>+</sup> reductases (FPR) primarily catalyze the transfer of reducing equivalents from NADPH to ferredoxin (or flavodoxin) to provide low potential reducing equivalents for the oxidoreductive metabolism. In addition, they can be implicated in regulating reactive oxygen species levels. Here we assess the functionality of FPR from B. ovis to understand its potential roles in the bacteria physiology. We prove that this FPR is active with the endogenous [2Fe-2S] Fdx ferredoxin, exhibiting a K<sub>M</sub><sup>Fdx</sup> in the low micromolar range. At the molecular level, this study provides with the first structures of an FPR at room temperature obtained by serial femtosecond crystallography, envisaging increase in flexibility at both the adenine nucleotide moiety of FAD and the C-terminal tail. The produced microcrystals are in addition suitable for future mix-and-inject time-resolved studies with the NADP<sup>+</sup>/H coenzyme either at synchrotrons or XFELs. Furthermore, the study also predicts the ability of FPR to simultaneously interact with Fdx and NADP<sup>+</sup>/H.
Authors
- Marta Martínez‐Júlvez (ORCID: https://orcid.org/0000-0001-9047-0046)
- Leland Bruce Gee (ORCID: https://orcid.org/0000-0002-5817-3997)
- Raymond G. Sierra (ORCID: https://orcid.org/0000-0002-6253-8282)
- Raphaël de Wijn (ORCID: https://orcid.org/0000-0002-8342-1008)
- Christina Schmidt (ORCID: https://orcid.org/0000-0002-6662-1294)
- Sabine Botha (ORCID: https://orcid.org/0000-0003-4362-7543)
- Peter Smyth
- Joana Valério (ORCID: https://orcid.org/0000-0001-5931-0925)
- Huijong Han (ORCID: https://orcid.org/0000-0002-1197-3014)
- Milagros Medina (ORCID: https://orcid.org/0000-0001-8743-0182)
- Joachim Schulz (ORCID: https://orcid.org/0000-0002-9310-8832)
- Christopher Kupitz (ORCID: https://orcid.org/0000-0002-7940-3042)
- S. Andrea Moreno (ORCID: https://orcid.org/0000-0002-5535-2762)
- E. V. Sobolev (ORCID: https://orcid.org/0000-0003-2478-5685)
- Stella Lisova (ORCID: https://orcid.org/0000-0002-6621-4285)
- Richard Bean (ORCID: https://orcid.org/0000-0001-8151-7439)
- Adam Round (ORCID: https://orcid.org/0000-0002-0723-8228)
- F. Trost (ORCID: https://orcid.org/0000-0001-5628-598X)
- E Juncheng
- Marcin Sikorski (ORCID: https://orcid.org/0009-0008-1243-4785)
- Katerina Dörner (ORCID: https://orcid.org/0000-0003-1072-5905)
- José M. Martín-García (ORCID: https://orcid.org/0000-0002-4558-3858)
- Marco Kloos (ORCID: https://orcid.org/0000-0002-8745-9740)
- Isabel Quereda‐Moraleda
- Valerio Mariani (ORCID: https://orcid.org/0000-0001-7611-5128)
- D. Melo
- Pamela Schleissner (ORCID: https://orcid.org/0000-0003-3855-9136)
- Celia Lozano-Vallhonrat
- María Buñuel-Escudero
Institutions
- Consejo Superior de Investigaciones Científicas (ES)
- Universidad de Zaragoza (ES)
- SLAC National Accelerator Laboratory (US)
- National Research Council (LK)
- Arizona State University (US)
- European X-Ray Free-Electron Laser (DE)
Publication Details
- Journal
- Archives of Biochemistry and Biophysics
- Published
- 2024-11-08
- DOI
- https://doi.org/10.1016/j.abb.2024.110204
- Citations
- 1
- Primary Topic
- Porphyrin Metabolism and Disorders
- Type
- article
- Field-Weighted Citation Impact
- 0.23
Funders
- Comunidad de Madrid
- European Commission
- Universidad de Zaragoza
- European Regional Development Fund
- Agencia Estatal de Investigación