000130875 001__ 130875
000130875 005__ 20240201151019.0
000130875 0247_ $$2doi$$a10.1016/j.bbabio.2019.148058
000130875 0248_ $$2sideral$$a114136
000130875 037__ $$aART-2019-114136
000130875 041__ $$aeng
000130875 100__ $$aPerez-Amigot, Daniel
000130875 245__ $$aTowards the competent conformation for catalysis in the ferredoxin-NADP(+) reductase from the Brucella ovis pathogen
000130875 260__ $$c2019
000130875 5203_ $$aBrucella ovis encodes a bacterial subclass 1 ferredoxin-NADP(H) reductase (BoFPR) that, by similarity with other FPRs, is expected either to deliver electrons from NADPH to the redox-based metabolism and/or to oxidize NADPH to regulate the soxRS regulon that protects bacteria against oxidative damage. Such potential roles for the pathogen survival under infection conditions make of interest to understand and to act on the BoFPR mechanism. Here, we investigate the NADP(+)/H interaction and NADPH oxidation by hydride transfer (HT) to BoFPR. Crystal structures of BoFPR in free and in complex with NADP(+) hardly differ. The latter shows binding of the NADP(+) adenosine moiety, while its redox-reactive nicotinamide protrudes towards the solvent. Nonetheless, pre-steady-state kinetics show formation of a charge-transfer complex (CTC-1) prior to the hydride transfer, as well as conversion of CTC-1 into a second charge-transfer complex (CTC-2) concomitantly with the HT event. Thus, during catalysis nicotinamide and flavin reacting rings stack. Kinetic data also identify the HT itself as the rate limiting step in the reduction of BoFPR by NADPH, as well as product release limiting the overall reaction. Using all-atom molecular dynamics simulations with a thermal effect approach we are able to visualise a potential transient catalytically competent interaction of the reacting rings. Simulations indicate that the architecture of the FAD folded conformation in BoFPR might be key in catalysis, pointing to its adenine as an element to orient the reactive atoms in conformations competent for HT.
000130875 536__ $$9info:eu-repo/grantAgreement/ES/DGA/E35-17R$$9info:eu-repo/grantAgreement/ES/MINECO-AEI-FEDER/BIO2016-75183-P
000130875 540__ $$9info:eu-repo/semantics/openAccess$$aAll rights reserved$$uhttp://www.europeana.eu/rights/rr-f/
000130875 590__ $$a3.465$$b2019
000130875 591__ $$aBIOPHYSICS$$b18 / 71 = 0.254$$c2019$$dQ2$$eT1
000130875 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b125 / 296 = 0.422$$c2019$$dQ2$$eT2
000130875 592__ $$a2.001$$b2019
000130875 593__ $$aBiochemistry$$c2019$$dQ1
000130875 593__ $$aCell Biology$$c2019$$dQ1
000130875 593__ $$aBiophysics$$c2019$$dQ1
000130875 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000130875 700__ $$0(orcid)0000-0001-9224-5854$$aTaleb, Víctor$$uUniversidad de Zaragoza
000130875 700__ $$aBoneta, Sergio
000130875 700__ $$0(orcid)0000-0002-6649-9153$$aAnoz-Carbonell, Ernesto$$uUniversidad de Zaragoza
000130875 700__ $$0(orcid)0000-0001-7202-4587$$aSebastian, María
000130875 700__ $$0(orcid)0000-0001-5702-4538$$aVelazquez-Campoy, Adrián$$uUniversidad de Zaragoza
000130875 700__ $$0(orcid)0000-0001-5823-7965$$aPolo, Víctor$$uUniversidad de Zaragoza
000130875 700__ $$0(orcid)0000-0001-9047-0046$$aMartinez-Julvez, Marta$$uUniversidad de Zaragoza
000130875 700__ $$0(orcid)0000-0001-8743-0182$$aMedina, Milagros$$uUniversidad de Zaragoza
000130875 7102_ $$12012$$2755$$aUniversidad de Zaragoza$$bDpto. Química Física$$cÁrea Química Física
000130875 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000130875 773__ $$g1860, 10 (2019), 148058 [12 pp.]$$pBiochim. biophys. acta, Bioenerg.$$tBIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS$$x0005-2728
000130875 8564_ $$s2279438$$uhttps://zaguan.unizar.es/record/130875/files/texto_completo.pdf$$yVersión publicada
000130875 8564_ $$s2404734$$uhttps://zaguan.unizar.es/record/130875/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000130875 909CO $$ooai:zaguan.unizar.es:130875$$particulos$$pdriver
000130875 951__ $$a2024-02-01-14:36:49
000130875 980__ $$aARTICLE