The structure of Enterococcus faecalis thymidylate synthase provides clues about folate bacterial metabolism

Drug resistance to therapeutic antibiotics poses a challenge to the identification of novel targets and drugs for the treatment of infectious diseases. Infections caused by Enterococcus faecalis are a major health problem. Thymidylate synthase (TS) from E. faecalis is a potential target for antibacterial therapy. We have determined the X-ray crystallographic structure of the E. faecalis thymidylate synthase (EfTS), obtained as a native binary complex composed of EfTS and 5-formyltetrahydrofolate (5-FTHF). The structure provide evidence that EfTS is a half-the-sites reactive enzyme as 5-FTHF is bound to two of the four independent subunits present in the crystal asymmetric unit. 5-FTHF is a metabolite of the one-carbon transfer reaction catalysed by 5-formyltetrahydrofolate cycloligase. Kinetic studies show that 5-FTHF is a weak inhibitor of EfTS, suggesting that the EfTS-5-FTHF complex may function as a source of folates and/or may regulate one-carbon metabolism. The structure represents the first example of endogenous 5-FTHF bound to a protein involved in folate metabolism.