000110777 001__ 110777
000110777 005__ 20230706131422.0
000110777 0247_ $$2doi$$a10.1002/iub.2576
000110777 0248_ $$2sideral$$a126033
000110777 037__ $$aART-2021-126033
000110777 041__ $$aeng
000110777 100__ $$aMoreno, Andrea
000110777 245__ $$aCofactors and pathogens: Flavin mononucleotide and flavin adenine dinucleotide (FAD) biosynthesis by the FAD synthase from Brucella ovis
000110777 260__ $$c2021
000110777 5060_ $$aAccess copy available to the general public$$fUnrestricted
000110777 5203_ $$aThe biosynthesis of the flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), cofactors used by 2% of proteins, occurs through the sequential action of two ubiquitous activities: a riboflavinkinase (RFK) that phosphorylates the riboflavin (RF) precursor to FMN, and a FMN:adenylyltransferase (FMNAT) that transforms FMN into FAD. In most mammals two different monofunctional enzymes have each of these activities, but in prokaryotes a single bifunctional enzyme, FAD synthase (FADS), holds them. Differential structural and functional traits for RFK and FMNAT catalysis between bacteria and mammals, as well as within the few bacterial FADSs so far characterized, has envisaged the potentiality of FADSs from pathogens as targets for the development of species-specific inhibitors. Here, we particularly characterize the FADS from the ovine pathogen Brucella ovis (BoFADS), causative agent of brucellosis. We show that BoFADS has RFK activity independently of the media redox status, but its FMNAT activity (in both forward and reverse senses) only occurs under strong reducing conditions. Moreover, kinetics for flavin and adenine nucleotides binding to the RFK site show that BoFADS binds preferentially the substrates of the RFK reaction over the products and that the adenine nucleotide must bind prior to flavin entrapment. These results, together with multiple sequence alignments and phylogenetic analysis, point to variability in the less conserved regions as contributing to the species-specific features in prokaryotic FADSs, including those from pathogens, that allow them to adopt alternative strategies in FMN and FAD biosynthesis and overall flavin homeostasis.
000110777 536__ $$9info:eu-repo/grantAgreement/ES/DGA-FEDER/E35-20R$$9info:eu-repo/grantAgreement/ES/MICINN-AEI/PID2019-103901GB-I00
000110777 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000110777 590__ $$a4.709$$b2021
000110777 592__ $$a0.893$$b2021
000110777 594__ $$a7.1$$b2021
000110777 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b125 / 297 = 0.421$$c2021$$dQ2$$eT2
000110777 593__ $$aBiochemistry$$c2021$$dQ2
000110777 591__ $$aCELL BIOLOGY$$b98 / 195 = 0.503$$c2021$$dQ3$$eT2
000110777 593__ $$aMolecular Biology$$c2021$$dQ2
000110777 593__ $$aGenetics$$c2021$$dQ2
000110777 593__ $$aCell Biology$$c2021$$dQ2
000110777 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000110777 700__ $$aTaleb, Víctor
000110777 700__ $$aSebastián, María
000110777 700__ $$aAnoz-Carbonell, Ernesto
000110777 700__ $$0(orcid)0000-0001-9047-0046$$aMartinez-Júlvez, Marta$$uUniversidad de Zaragoza
000110777 700__ $$0(orcid)0000-0001-8743-0182$$aMedina, Milagros$$uUniversidad de Zaragoza
000110777 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000110777 773__ $$g74, 7 (2021), 655-671$$pIUBMB life$$tIUBMB LIFE$$x1521-6543
000110777 8564_ $$s7002769$$uhttps://zaguan.unizar.es/record/110777/files/texto_completo.pdf$$yVersión publicada
000110777 8564_ $$s2459879$$uhttps://zaguan.unizar.es/record/110777/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000110777 909CO $$ooai:zaguan.unizar.es:110777$$particulos$$pdriver
000110777 951__ $$a2023-07-06-12:21:12
000110777 980__ $$aARTICLE