148928 20260108074821.0 doi 10.1016/j.abb.2025.110288 sideral 142107 ART-2025-142107 eng (orcid)0000-0001-7947-8813 Minjárez-Sáenz, Martha Structural and functional insights into UDP-N-acetylglucosamine-enolpyruvate reductase (MurB) from Brucella ovis 2025 The peptidoglycan biosynthetic pathway involves a series of enzymatic reactions in which UDP-N-acetylglucosamine-enolpyruvate reductase (MurB) plays a crucial role in catalyzing the conversion of UDP-N-acetylglucosamine-enolpyruvate (UNAGEP) to UDP-N-acetylmuramic acid. This reaction relies on NADPH and FAD and, since MurB is not found in eukaryotes, it is an attractive target for the development of antimicrobials. MurB from Brucella ovis, the causative agent of brucellosis in sheep, is characterized here. The FAD cofactor in MurB of B. ovis is reduced to the hydroquinone state without semiquinone stabilization with an estimated Eox/hq of −260 mV. MurB from B. ovis catalyzes the oxidation of NADPH in a slow process that is positively influenced by the presence of the second product, UNAGEP. The crystallographic structure of the MurBox:UNAGEP complex confirms its folding into three domains and the binding of UNAGEP, positioning its enolpyruvyl group for hydride transfer from FAD. MurB shows a complex thermal unfolding pathway that is influenced by UNAGEP and NADP+, confirming its ability to bind both molecules. Molecular dynamics (MD) simulations predict that the nicotinamide of NADP+ is more stable at the active site than the enolpyruvyl of UNAGEP, and suggests that MurB can simultaneously accommodate NADPH and UNAGEP in the substrate channel, increasing overall protein-ligand flexibility. Sequence and evolutionary analyses show that MurB from B. ovis conserves all motifs predicted to be involved in catalysis within the Type IIa family. info:eu-repo/grantAgreement/ES/DGA/E35-23R info:eu-repo/grantAgreement/ES/DGA/LMP13_21 info:eu-repo/grantAgreement/ES/MICINN/PID2022-136369NB-I00 info:eu-repo/semantics/openAccess All rights reserved http://www.europeana.eu/rights/rr-f/ info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Rivero, Maribel Universidad de Zaragoza Correa-Pérez, Víctor Universidad de Zaragoza Boneta, Sergio Universidad de Zaragoza Suárez, Paula (orcid)0000-0001-5823-7965 Polo, Víctor Universidad de Zaragoza Sadeghi, Sheila J. (orcid)0000-0003-3608-4720 Yruela, Inmaculada (orcid)0000-0001-9047-0046 Martínez-Júlvez, Marta Universidad de Zaragoza (orcid)0000-0001-8743-0182 Medina, Milagros Universidad de Zaragoza 2012 755 Universidad de Zaragoza Dpto. Química Física Área Química Física 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 765 (2025), 110288 [19 pp.] Arch. biochem. biophys. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 0003-9861 12381399 http://zaguan.unizar.es/record/148928/files/texto_completo.pdf Versión publicada info:eu-repo/date/embargoEnd/2026-01-04 2477272 http://zaguan.unizar.es/record/148928/files/texto_completo.jpg?subformat=icon icon Versión publicada info:eu-repo/date/embargoEnd/2026-01-04 oai:zaguan.unizar.es:148928 articulos driver 2026-01-07-18:55:46 ARTICLE