TY - JOUR
T1 - The molecular basis of pyrazinamide activity on Mycobacterium tuberculosis PanD
AU - Sun, Qingan
AU - Li, Xiaojun
AU - Perez, Lisa M.
AU - Shi, Wanliang
AU - Zhang, Ying
AU - Sacchettini, James C.
N1 - Funding Information:
Funding for this work came from the TB Structural Genomics Consortium, P01AI095208 from NIAID, NIH (J.C.S.), grant A-0015 from the Welch Foundation (J.C.S.), and a grant from the Chancellor Research Initiative-Texas A&M System (J.C.S.). Results shown in this report are derived from work performed at Argonne National Laboratory, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory. Data were collected from the Structural Biology Center (SBC) at the Advanced Photon Source (Argonne National Laboratory). SBC-CAT is operated by U. Chicago Argonne, LLC, for the U.S. Department of Energy, Office of Biological and Environmental Research under contract DE-AC02-06CH11357. We thank the staff at beamline 19-ID and 23-ID for assistance with data collection. We thank Dr. Steve Lockless for advice on ITC and reading the manuscript, Dr. Matthews Benning for advice on X-ray data processing with Proteum (Bruker), and Tracey Musa for editing the manuscript.
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - 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.
AB - 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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U2 - 10.1038/s41467-019-14238-3
DO - 10.1038/s41467-019-14238-3
M3 - Article
C2 - 31953389
AN - SCOPUS:85078061351
SN - 2041-1723
VL - 11
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 339
ER -