Role of pncA gene mutations W68R and W68G in pyrazinamide resistance

• Published in: Journal of cellular biochemistry 2018-03, Vol.119 (3), p.2567-2578
• Main Authors: Aggarwal, Mansi, Singh, Aditi, Grover, Sonam, Pandey, Bharati, Kumari, Anchala, Grover, Abhinav
• Format: Article
• Language: English
• Subjects: Affinity; Bacilli; Binding sites; Comparative analysis; Complementarity; Docking; Drug resistance; Drugs; Molecular dynamics; molecular dynamics simulation; Mutation; Mycobacterium tuberculosis; PncA gene; Proteins; pyrazinamidase; Pyrazinamide; PZA resistance; Tuberculosis
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/jcb.26420

Mycobacterium tuberculosis (Mtb) resistance toward anti‐tuberculosis drugs is a widespread problem. Pyrazinamide (PZA) is a first line antitubercular drug that kills semi‐dormant bacilli when converted into its activated form, that is, pyrazinoic acid (POA) by Pyrazinamidase (PZase) enzyme coded by pncA gene. In this study, we conducted several analyses on native and mutant structures (W68R, W68G) of PZase before and after docking with the PZA drug to explore the molecular mechanism behind PZA resistance caused due to pncA mutations. Structural changes caused by mutations were studied with respect to their effects on functionality of protein. Docking was performed to analyze the protein‐drug binding and comparative analysis was done to observe how the mutations affect drug binding affinity and binding site on protein. Native PZase protein was observed to have the maximum binding affinity in terms of docking score as well as shape complementarity in comparison to the mutant forms. Molecular dynamics simulation analyses showed that mutation in the 68th residue of protein results in a structural change at its active site which further affects the biological function of protein, that is, conversion of PZA to POA. Mutations in the protein thereby led to PZA resistance in the bacterium due to the inefficient binding. In this study, we conducted several analyses on native and mutant structures (W68R, W68G) of Pyrazinamidase (PZase) before and after docking with the PZA drug to explore the mechanism behind PZA resistance. Molecular dynamics simulation analyses showed that mutation in the 68th residue of protein results in a structural change at its active site which further affects the biological function of protein, that is, to act as an enzyme and convert PZA into pyrazinoic acid (POA). The mutations in the protein thereby led to PZA resistance in the bacterium due to the inefficient binding.