The molecular basis of pyrazinamide activity on Mycobacterium tuberculosis PanD.
Qingan Sun, Xiaojun Li, Lisa M Perez, Wanliang Shi, Ying Zhang, James C Sacchettini
Author Information
Qingan Sun: Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX, USA. ORCID
Xiaojun Li: Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX, USA.
Lisa M Perez: Laboratory for Molecular Simulation, Texas A&M University, College Station, TX, USA. ORCID
Wanliang Shi: Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, 21205, USA.
Ying Zhang: Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, 21205, USA.
James C Sacchettini: Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX, USA. sacchett@tamu.edu.
Pyrazinamide has been a mainstay in the multidrug regimens used to treat tuberculosis. It is active against the persistent, non-replicating mycobacteria responsible for the protracted therapy required to cure tuberculosis. Pyrazinamide is a pro-drug that is converted into pyrazinoic acid (POA) by pyrazinamidase, however, the exact target of the drug has been difficult to determine. Here we show the enzyme PanD binds POA in its active site in a manner consistent with competitive inhibition. The active site is not directly accessible to the inhibitor, suggesting the protein must undergo a conformational change to bind the inhibitor. This is consistent with the slow binding kinetics we determined for POA. Drug-resistant mutations cluster near loops that lay on top of the active site. These resistant mutants show reduced affinity and residence time of POA consistent with a model where resistance occurs by destabilizing the closed conformation of the active site.
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