Discovery of Selective Small-Molecule Activators of a Bacterial Glycoside Hydrolase.

摘要: Fragment-based approaches are used routinely to discover enzyme inhibitors as cellular tools and potential therapeutic agents. There have been few reports, however, of the discovery small-molecule activators. Herein, we describe characterization activators a glycoside hydrolase (a bacterial O-GlcNAc hydrolase). A ligand-observed NMR screen library commercially available fragments identified an activator which yielded approximate 90 % increase in kcat/KM values (kcat=catalytic rate constant; KM=Michaelis constant). This compound binds close proximity catalytic center. Evolution initial hits led improved compounds that behave nonessential effecting both KM and Vmax values (Vmax=maximum reaction). The appear stabilize active “closed” form enzyme. Such could offer orthogonal alternative for perturbation activity in vivo, also be activation many industrial processes.