Cofactors and pathogens: Flavin mononucleotide and flavin adenine dinucleotide (FAD) biosynthesis by the FAD synthase from Brucella ovis

Moreno, Andrea ; Taleb, Víctor ; Sebastián, María ; Anoz-Carbonell, Ernesto ; Martinez-Júlvez, Marta (Universidad de Zaragoza) ; Medina, Milagros (Universidad de Zaragoza)

Resumen: The 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.

Idioma: Inglés

DOI: 10.1002/iub.2576

Año: 2021

Publicado en: IUBMB LIFE 74, 7 (2021), 655-671
ISSN: 1521-6543

Factor impacto JCR: 4.709 (2021)

Categ. JCR: BIOCHEMISTRY & MOLECULAR BIOLOGY rank: 125 / 297 = 0.421 (2021) - Q2 - T2
Categ. JCR: CELL BIOLOGY rank: 98 / 195 = 0.503 (2021) - Q3 - T2

Factor impacto CITESCORE: 7.1 - Biochemistry, Genetics and Molecular Biology (Q2)

Factor impacto SCIMAGO: 0.893 - Biochemistry (Q2) - Molecular Biology (Q2) - Genetics (Q2) - Cell Biology (Q2)

Financiación: info:eu-repo/grantAgreement/ES/DGA-FEDER/E35-20R
Financiación: info:eu-repo/grantAgreement/ES/MICINN-AEI/PID2019-103901GB-I00

Tipo y forma: Article (Published version)

Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)