Investigating the Roles of Active Site Residues in Mycobacterium tuberculosis Indole-3-glycerol Phosphate Synthase, a Potential Target for Antitubercular Agents.

Mycobacterium tuberculosis drug resistance is emerging and new drug targets are needed. Tryptophan biosynthesis is necessary for M. tuberculosis replication and virulence. Indole-3-glycerol phosphate synthase (IGPS) catalyzes a step in M. tuberculosis tryptophan biosynthesis and has been suggested as a potential anti-infective target, but our understanding of this enzyme is limited. To aid in inhibitor design and gain a greater mechanistic picture of this enzyme, there is a need to understand the roles of active site amino acids in ligand binding and catalysis. In this work, we explored the roles of conserved active site amino acids Glu57, Lys59, Lys119, Glu168, and Glu219. Mutation of each to Ala results in loss of all detectable activity. The Glu57Gln, Lys59Arg, Lys119Arg, Glu168Gln, and Glu219Asp mutations result in large activity losses, while Glu219Gln has enhanced activity. Analysis of the enzymatic data yields the following main conclusions: (A) Lys119 is the likely catalytic acid in the CdRP ring closure step. (B) Glu168 stabilizes a charged reaction intermediate and may also be the catalytic base. (C) Glu57, Glu219, and Lys119 form a closely arranged triad in which Glu57 and Glu219 modulate the pKa of Lys119, and thus overall activity. This increased understanding of inter- and intramolecular interactions and demonstration of the highly coordinated nature of the M. tuberculosis IGPS active site provide new mechanistic information and guidance for future work with this potential new drug target.

Degradation Mechanism Study and Safety Hazard Analysis of Overdischarge on Commercialized Lithium-ion Batteries.

With the continuous improvement of the energy density of traction batteries for electric vehicles, the safety of batteries over their entire lifecycle has become the most critical issue in the development of electric vehicles. Abuse of electricity encountered in the application of batteries has a great impact on the safety of traction batteries. In this study, focused on the overdischarge phenomenon that is most likely to be encountered in the practical use of electric vehicles and grid storage, the impact of overdischarge on battery performance degradation is analyzed by neutron imaging technology and its safety hazards is systematically explored, combined with multimethods including electrochemical analysis and structural characterization. Results reveal the deterioration of the internal structure of traction batteries due to the overdischarge behavior and play a guiding role in the testing and evaluation of the safety of traction batteries.