Structural study and thermodynamic characterization of inhibitor binding to lumazine synthase from Bacillus anthracis

The crystal structure of lumazine synthase from Bacillus anthracis was solved by molecular replacement and refined to R (cryst) = 23.7% (R (free) = 28.4%) at a resolution of 3.5 Å. The structure reveals the icosahedral symmetry of the enzyme and specific features of the active site that are unique in comparison with previously determined orthologues. The application of isothermal titration calorimetry in combination with enzyme kinetics showed that three designed pyrimidine derivatives bind to lumazine synthase with micromolar dissociation constants and competitively inhibit the catalytic reaction. Structure-based modelling suggested the binding modes of the inhibitors in the active site and allowed an estimation of the possible contacts formed upon binding. The results provide a structural framework for the design of antibiotics active against B. anthracis.