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oai:figshare.com:article/34036855 2026-09-30T20:40:02Z category_4 category_7 category_14 category_19 category_21 category_734 category_931 category_64 category_132 category_134 portal_87 item_type_6 month_year_09_2026

Computational and biophysical validation of tobramycin targeting Mycobacterium MetE(Rv1133c): implications for anti-tuberculosis activity & rifampicin synergy Kunal Malik (25146118) Kashish Choudhary (25146121) Waseem Ali (6726656) Sana Tanweer (19199245) Rahul Sharma (1458472) Rahul Verma (1448428) Sana Jamily (25146124) Shivani A. Muthu (9333165) Meetu Agarwal (14755869) Abhinav Grover (185963) Laishram Rajendrakumar Singh (9464246) Basir Ahmad (290508) Sonam Grover (849429) Biochemistry Medicine Molecular Biology Pharmacology Biotechnology Biological Sciences not elsewhere classified Information Systems not elsewhere classified Cancer Infectious Diseases Virology Drug repurposing methionine synthase MetE mycobacterium tuberculosis tuberculosis The global escalation of multidrug-resistant tuberculosis (MDR-TB) demands identification of novel therapeutic targets. Here, we establish Vitamin B12-independent methionine synthase E ‘MetE’ (Rv1133c) as a crucial drug target and validate tobramycin as its inhibitor. Through a multi-faceted approach, including virtual screening and MD simulations, we identified interactions between the tobramycin-MetE complex, which were experimentally validated. Through Isothermal calorimetry we found tobramycin binds to MetE with high affinity (Ka⁓10−8M). Subsequent biophysical characterization revealed an interesting mechanism of action: tobramycin binding induces conformational perturbations of the MetE enzyme, as confirmed by changes in intrinsic fluorescence, denaturation patterns and circular dichroism. Under In-vitro conditions tobramycin demonstrated potent anti-mycobacterial activity at concentrations nontoxic to human macrophage cells. We hypothesize that this inhibition involves classical 30S ribosomal binding and MetE disruption. Most significantly, we discovered a powerful synergistic effect between tobramycin and the first-line anti-TB drug, rifampicin. Together, these findings establish a robust preclinical foundation for repurposing tobramycin against management of tuberculosis. Flowchart illustrating tuberculosis treatment strategies, including DOTS, drug repurposing, and virtual screening.The image is a flowchart summarizing tuberculosis (TB) treatment strategies. At the top, "infected lungs" and an inset depicting active TB in alveolar macrophages highlight the infection. Below, treatment paths show "DOTS Treatment" with limits for multidrug-resistant (MDR) and extensively drug-resistant TB (XDR-TB) crossed out, while "Drug Repurposing" indicates potential benefits for these strains. Arrows lead to an "FDA approved drug library" connected to "Virtual Screening" that produces "Tobramycin" and progresses to "Biophysical Characterization of MetE-Tobramycin". Results include "Synergy with Rifampicin" and "In-vitro anti-mycobacterial activity". For the first time we are establishing MetE as a drug target in M.tb which is absent in humans, making it crucial.As a part of drug repurposing, for the first time we are validating tobramycin as the first-known inhibitor of MetE.We propose that tobramycin’s anti-mycobacterial efficacy is driven by a combination of its classical role in inhibiting protein synthesis and disruption of MetE-mediated methionine biosynthesis.We demonstrated synergistic interaction between tobramycin and rifampicin. This combinatory therapy drastically reduces the required concentrations of both drugs, offering a novel anti-TB strategy with the potential to lower costs, shorten treatment, and combat MDR-TB. For the first time we are establishing MetE as a drug target in M.tb which is absent in humans, making it crucial. As a part of drug repurposing, for the first time we are validating tobramycin as the first-known inhibitor of MetE. We propose that tobramycin’s anti-mycobacterial efficacy is driven by a combination of its classical role in inhibiting protein synthesis and disruption of MetE-mediated methionine biosynthesis. We demonstrated synergistic interaction between tobramycin and rifampicin. This combinatory therapy drastically reduces the required concentrations of both drugs, offering a novel anti-TB strategy with the potential to lower costs, shorten treatment, and combat MDR-TB. 2026-09-30T20:40:02Z Text Journal contribution 10.6084/m9.figshare.34036855.v1 https://figshare.com/articles/journal_contribution/Computational_and_biophysical_validation_of_tobramycin_targeting_i_Mycobacterium_i_MetE_Rv1133c_implications_for_anti-tuberculosis_activity_rifampicin_synergy/34036855 CC BY 4.0