posted on 2023-02-02, 22:35authored byMG Peverelli, Tatiana Soares-da-Costa, N Kirby, Matthew Perugini
Diaminopimelate decarboxylase (DAPDC) catalyzes the final step in the diaminopimelate biosynthesis pathway of bacteria. The product of the reaction is the essential amino acid L-lysine, which is an important precursor for the synthesis of the peptidoglycan cell wall, housekeeping proteins, and virulence factors of bacteria. Accordingly, the enzymeisapromising antibacterial target. Previous structural studies demonstrate that DAPDC exists as monomers, dimers, and tetramers in the crystal state. However, the active oligomeric form has not yet been determined. We show using analytical ultracentrifugation, small angle x-ray scattering, and enzyme kinetic analyses in solution that the active form of DAPDC from Bacillus anthracis, Escherichia coli, Mycobacterium tuberculosis, and Vibrio cholerae is a dimer. The importance of dimerization was probed further by generating dimerization interface mutants (N381A and R385A) of V. cholerae DAPDC. Our studies indicate that N381A and R385A are significantly attenuated in catalytic activity, thus confirming that dimerization of DAPDC is essential for function. These findings provide scope for the development of new antibacterial agents that prevent DAPDC dimerization.
Funding
This work was supported in part by Australian Research Council (ARC) Discovery Project DP150103313. The authors declare that they have no conflicts of interest with the contents of this article.Supported by National Health and Medical Research Council of Australia Grant APP1091976 for fellowship support.
History
Publication Date
2016-04-29
Journal
Journal of Biological Chemistry
Volume
291
Issue
18
Pagination
11p. (p. 9785-9795)
Publisher
American Society for Biochemistry and Molecular Biology