Publication Type:

Journal Article

Source:

International Journal of Antimicrobial Agents, Volume 37, Number 4, p.368-372 (2011)

URL:

http://www.scopus.com/inward/record.url?eid=2-s2.0-79952815955&partnerID=40&md5=558539d5faf23ea397b65b7c89e1585f

Keywords:

antibiotic sensitivity, Antitubercular Agents, article, bacterial gene, Bacterial Proteins, bacterial strain, Base Sequence, bioinformatics, catalase, Computational Biology, controlled study, DNA Primers, Drug Resistance, enzyme active site, enzyme binding, enzyme conformation, enzyme stability, enzyme structure, ethambutol, gene mutation, gene targeting, glutamine, histidine, India, isoniazid, katG gene, KatG protein, methionine, Microbial, Microbial Sensitivity Tests, Models, Molecular, molecular docking, molecular dynamics, Molecular Dynamics Simulation, molecular mechanics, multidrug resistance, Multiple, Mutation, mutational analysis, Mycobacterium tuberculosis, nonhuman, nucleotide sequence, polymerase chain reaction, prediction, priority journal, pyrazinamide, rifampicin, serine, streptomycin, threonine, wild type

Abstract:

This study reports on the structural basis of drug resistance targeting the katG gene in a multidrug-resistant Mycobacterium tuberculosis (MDR-TB) strain with two novel mutations (His276Met and Gln295His) in addition to the most commonly reported mutation (Ser315Thr). A structural bioinformatics approach was used to predict the structure of the mutant KatG enzyme (MT). Subsequent molecular dynamics and docking studies were performed to explain the mechanism of isoniazid (INH) resistance. The results show significant conformational changes in the structure of MT leading to a change in INH binding residues at the active site, with a significant increase in the inhibition constant (Ki) of 5.67 μm in the mutant KatG-isoniazid complex (MT-INH) compared with the wild-type KatG-isoniazid complex (WT-INH). In the case of molecular dynamics studies, root mean square deviation (RMSD) analysis of the protein backbone in simulated biological conditions revealed an unstable trajectory with higher deviations in MT throughout the simulation process (1 ns). Moreover, root mean square fluctuation (RMSF) analysis revealed an overall increase in residual fluctuations in MT compared with the wild-type KatG enzyme (WT), whilst the INH binding residues of MT showed a decreased fluctuation that can be observed as peak deviations. Hence, the present study suggests that His276Met, Gln295His and Ser315Thr mutations targeting the katG gene result in decreased stability and flexibility of the protein at INH binding residues leading to impaired enzyme function. © 2010 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

Notes:

cited By (since 1996)2

Cite this Research Publication

Ga Ramasubban, Therese, K. La, Vetrivel, Ub, Sivashanmugam, Mb, Rajan, Pc, Sridhar, Rd, Madhavan, H. Na, and Meenakshi, Ne, “Detection of novel coupled mutations in the katG gene (His276Met, Gln295His and Ser315Thr) in a multidrug-resistant Mycobacterium tuberculosis strain from Chennai, India, and insight into the molecular mechanism of isoniazid resistance using structural bi”, International Journal of Antimicrobial Agents, vol. 37, pp. 368-372, 2011.