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Please use this identifier to cite or link to this item: http://repository.iitr.ac.in/handle/123456789/20245
Title: Variations in the SDN Loop of Class A Beta-Lactamases: A Study of the Molecular Mechanism of BlaC (Mycobacterium tuberculosis) to Alter the Stability and Catalytic Activity Towards Antibiotic Resistance of MBIs
Authors: Bhattacharya S.
Junghare V.
Pandey N.K.
Baidya S.
Agarwal H.
Das N.
Banerjee A.
Ghosh D.
Roy, Partha
Patra H.K.
Hazra, Saugata
Published in: Frontiers in Microbiology
Abstract: The emergence of multidrug-resistant (MDR) and extensively drug-resistant (XDR) tuberculosis calls for an immediate search for novel treatment strategies. Recently, BlaC, the principal beta-lactamase of Mycobacterium tuberculosis, was recognized as a potential therapeutic target. BlaC belongs to Ambler class A, which is generally susceptible to the beta-lactamase inhibitors currently used in clinics: tazobactam, sulbactam, and clavulanate. Alterations at Ser130 in conserved SDN loop confer resistance to mechanism-based inhibitors (MBIs) commonly observed in various clinical isolates. The absence of clinical evidence of S130G conversion in M. tuberculosis draws our attention to build laboratory mutants of S130G and S130A of BlaC. The study involving steady state, inhibition kinetics, and fluorescence microscopy shows the emergence of resistance against MBIs to the mutants expressing S130G and S130A. To understand the molecular reasoning behind the unavailability of such mutation in real life, we have used circular dichroism (CD) spectroscopy, differential scanning calorimetry (DSC), molecular dynamics (MD) simulation, and stability-based enzyme activity to compare the stability and dynamic behaviors of native and S130G/A mutant form of BlaC. A significant decrease in melting temperature (BlaC TM 60°C, S130A TM 50°C, and S130G TM 45°C), kinetic instability at higher temperature, and comparative dynamic instability correlate the fact that resistance to beta-lactam/beta-lactamase inhibitor combinations will likely not arise from the structural alteration of BlaC, therefore establishing confidence that this therapeutic modality can be potentially applied as a part of a successful treatment regimen against M. tuberculosis. © Copyright © 2021 Bhattacharya, Junghare, Pandey, Baidya, Agarwal, Das, Banerjee, Ghosh, Roy, Patra and Hazra.
Citation: Frontiers in Microbiology, 12
URI: https://doi.org/10.3389/fmicb.2021.710291
http://repository.iitr.ac.in/handle/123456789/20245
Issue Date: 2021
Publisher: Frontiers Media S.A.
Keywords: antimicrobial resistance (AMR)
combinatorial therapeutics
conserved loop
extensively drug-resistant (XDR)
MD simulation
mechanism-based inhibitors (MBI)
multidrug-resistant (MDR)
Mycobacterium tuberculosis
ISSN: 1664302X
Author Scopus IDs: 57202078739
57200569168
57213174613
57304488000
57304183300
57209505172
57216363128
9941456000
35509207200
14067825400
15837197000
Author Affiliations: Bhattacharya, S., Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, India
Junghare, V., Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, India
Pandey, N.K., Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, India
Baidya, S., Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, India
Agarwal, H., Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, India
Das, N., Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, India
Banerjee, A., Biochemistry and BIotechnology Area, Material Resource Efficiency Division, CSIR-Indian Institute of Petroleum, Dehradun, India, Academy of Scientific and Innovative Research, Ghaziabad, India
Ghosh, D., Biochemistry and BIotechnology Area, Material Resource Efficiency Division, CSIR-Indian Institute of Petroleum, Dehradun, India, Academy of Scientific and Innovative Research, Ghaziabad, India
Roy, P., Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, India
Patra, H.K., Department of Surgical Biotechnology, University College London, London, United Kingdom
Hazra, S., Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee, India, Centre for Nanotechnology, Indian Institute of Technology Roorkee, Roorkee, India
Funding Details: A part of the work was conducted in the Department of Biotechnology central facility IIT Roorkee, ICC IIT Roorkee, and Bioinformatics Center, Department of Biotechnology, IIT Roorkee. Financial support through ICMR (Indian Council of Medical Research) sanction number BIC/12(15)/2014, FIG (IITR) sanction number FIG 100671/14, SERB (Science and Engineering Research Board, India) sanction number YSS/2014/000492, and SPARC (Scheme for Promotion of Academic and Research Collaboration, Minister of Human Resource development, India) sanction number SPARC/2018-2019/8/SL(IN) to SH are gratefully acknowledged. SPARC/2018-2019/8/SL; Indian Council of Medical Research, ICMR: BIC/12(15)/2014, FIG 100671/14; Science and Engineering Research Board, SERB: YSS/2014/000492
Corresponding Author: Hazra, S.; Department of Biosciences and Bioengineering, India; email: saugata.iitk@gmail.com
Appears in Collections:Journal Publications [BT]

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