The FAD synthetase from the human pathogen Streptococcus pneumoniae: a bifunctional enzyme exhibiting activity-dependent redox requirements

M. Sebastián, E. Lira-Navarrete, A. Serrano, C. Marcuello, A. Velázquez-Campoy, A. Lostao, R. Hurtado-Guerrero, M. Medina & M. Martínez-Júlvez. Scientific Reports 7, 7609, 2017

Prokaryotic bifunctional FAD synthetases (FADSs) catalyze the biosynthesis of FMN and FAD, whereas in eukaryotes two enzymes are required for the same purpose. FMN and FAD are key cofactors to maintain the flavoproteome homeostasis in all type of organisms. Here we shed light to the properties of the hitherto unstudied bacterial FADS from the human pathogen Streptococcus pneumoniae (SpnFADS). As other members of the family, SpnFADS catalyzes the three typical activities of prokaryotic FADSs: riboflavin kinase (RFK), ATP:FMN:adenylyltransferase (FMNAT), and FAD pyrophosphorylase (FADpp). However, several SpnFADS biophysical properties differ from those of other family members. In particular; i) the RFK activity is not inhibited by the riboflavin (RF) substrate, ii) the FMNAT and FADSpp activities require flavin substrates in the reduced state, iii) binding of adenine nucleotide ligands is required for the binding of flavinic substrates/products and iv) the monomer is the preferred state. Collectively, our results add interesting mechanistic differences among the few prokaryotic bifunctional FADSs already characterized, which might reflect the adaptation of the enzyme to relatively different environments. In a health point of view, differences among FADS family members provide us with a framework to design selective compounds targeting these enzymes for the treatment of diverse infectious diseases. Prokaryotic FAD synthetases (FADSs) are bifunctional and bimodular enzymes that first catalyze the synthesis of flavin mononucleotide (FMN) from riboflavin (RF, vitamin B 2) through an ATP:riboflavin kinase activity (RFK, EC 2.7.1.26) and subsequently the adenylylation of FMN to produce flavin adenine dinucleotide (FAD) using an ATP:FMN:adenylyltransferase activity (FMNAT, EC 2.7.7.2) 1–3. For some of them, the FMNAT activity has also been shown to be reversible, exhibiting therefore a FAD pyrophosphorylase activity (FADpp) 4, 5. In mammals and yeasts two independent enzymes catalyze these reactions 6–9. Eukaryotic RFKs exhibit structural homol-ogy with the C-terminal module of prokaryotic bifunctional FADSs 10–12. On the contrary eukaryotic FMNATs do not exhibit similarity to the prokaryotic N-terminal modules, which belong to the nucleotidyltransferase.

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