Examining the role of protein structural dynamics in drug resistance in Mycobacterium tuberculosis

Shaw, Daniel; Hill, Rachel; Simpson, Niall; Husseini, Fouad; Robb, Kirsty; Greetham, Gregory M; Towrie, Michael; Parker, Anthony William; Robinson, David; Hirst, Jonathan; Hoskisson, Paul A; Hunt, Neil

Abstract

Antimicrobial resistance represents a growing global health problem. The emergence of novel resistance mechanisms necessitates the development of alternative approaches to investigate the molecular fundamentals of resistance, leading ultimately to new strategies for counteracting them. To gain deeper insight into antibiotic-target interactions, the binding of the frontline anti-tuberculosis drug isoniazid (INH) to a target enzyme, InhA, from Mycobacterium tuberculosis was studied using ultrafast two-dimensional infrared (2D-IR) spectroscopy and molecular simulations. Comparing wild-type InhA with a series of single point mutations, it was found that binding of the INH-NAD inhibitor to susceptible forms of the enzyme caused increased vibrational coupling between residues located in the Rossmann fold co-factor binding site of InhA, reducing dynamic fluctuations. The effect correlated with biochemical assay data, being markedly reduced in the INH-resistant S94A mutant and absent in the biochemically-inactive P193A control. Molecular dynamics simulations and calculations of inter-residue couplings indicate that the changes in coupling and dynamics are not localised to the co-factor binding site, but permeate much of the protein. We thus propose that the resistant S94A mutation circumvents subtle changes in global structural dynamics caused by INH upon binding to the wild-type enzyme that may impact upon the formation of important protein-protein complexes in the fatty acid synthase pathway of M. tuberculosis.

Citation

Shaw, D., Hill, R., Simpson, N., Husseini, F., Robb, K., Greetham, G. M., Towrie, M., Parker, A. W., Robinson, D., Hirst, J., Hoskisson, P. A., & Hunt, N. (in press). Examining the role of protein structural dynamics in drug resistance in Mycobacterium tuberculosis. Chemical Science, 8, https://doi.org/10.1039/C7SC03336B

• Journal Article Type: Article
• Acceptance Date: Oct 16, 2017
• Online Publication Date: Oct 16, 2017
• Deposit Date: Oct 18, 2017
• Publicly Available Date: Oct 18, 2017
• Journal: Chemical Science
• Print ISSN: 2041-6520
• Electronic ISSN: 2041-6539
• Publisher: Royal Society of Chemistry
• Peer Reviewed: Peer Reviewed
• Volume: 8
• DOI: https://doi.org/10.1039/C7SC03336B
• Public URL: https://nottingham-repository.worktribe.com/output/888167
• Publisher URL: http://pubs.rsc.org/en/Content/ArticleLanding/2017/SC/C7SC03336B#!divAbstract
• Contract Date: Oct 18, 2017