Disruption of key NADH-binding pocket residues of the Mycobacterium tuberculosis InhA affects DD-CoA binding ability

Abstract Tuberculosis (TB) is a global health problem that affects over 10 million people. There is an urgent need to develop novel antimicrobial therapies to combat TB. To achieve this, a thorough understanding of key validated drug targets is required. The enoyl reductase InhA, responsible for syn...

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Autores principales:	Daniel J. Shaw, Kirsty Robb, Beatrice V. Vetter, Madeline Tong, Virginie Molle, Neil T. Hunt, Paul A. Hoskisson
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/969f64fbcfb949d18b2dcc5f36be6f93
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spelling	oai:doaj.org-article:969f64fbcfb949d18b2dcc5f36be6f932021-12-02T15:06:08ZDisruption of key NADH-binding pocket residues of the Mycobacterium tuberculosis InhA affects DD-CoA binding ability10.1038/s41598-017-05042-42045-2322https://doaj.org/article/969f64fbcfb949d18b2dcc5f36be6f932017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05042-4https://doaj.org/toc/2045-2322Abstract Tuberculosis (TB) is a global health problem that affects over 10 million people. There is an urgent need to develop novel antimicrobial therapies to combat TB. To achieve this, a thorough understanding of key validated drug targets is required. The enoyl reductase InhA, responsible for synthesis of essential mycolic acids in the mycobacterial cell wall, is the target for the frontline anti-TB drug isoniazid. To better understand the activity of this protein a series of mutants, targeted to the NADH co-factor binding pocket were created. Residues P193 and W222 comprise a series of hydrophobic residues surrounding the cofactor binding site and mutation of both residues negatively affect InhA function. Construction of an M155A mutant of InhA results in increased affinity for NADH and DD-CoA turnover but with a reduction in Vmax for DD-CoA, impairing overall activity. This suggests that NADH-binding geometry of InhA likely permits long-range interactions between residues in the NADH-binding pocket to facilitate substrate turnover in the DD-CoA binding region of the protein. Understanding the precise details of substrate binding and turnover in InhA and how this may affect protein-protein interactions may facilitate the development of improved inhibitors enabling the development of novel anti-TB drugs.Daniel J. ShawKirsty RobbBeatrice V. VetterMadeline TongVirginie MolleNeil T. HuntPaul A. HoskissonNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-7 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Daniel J. Shaw Kirsty Robb Beatrice V. Vetter Madeline Tong Virginie Molle Neil T. Hunt Paul A. Hoskisson Disruption of key NADH-binding pocket residues of the Mycobacterium tuberculosis InhA affects DD-CoA binding ability
description	Abstract Tuberculosis (TB) is a global health problem that affects over 10 million people. There is an urgent need to develop novel antimicrobial therapies to combat TB. To achieve this, a thorough understanding of key validated drug targets is required. The enoyl reductase InhA, responsible for synthesis of essential mycolic acids in the mycobacterial cell wall, is the target for the frontline anti-TB drug isoniazid. To better understand the activity of this protein a series of mutants, targeted to the NADH co-factor binding pocket were created. Residues P193 and W222 comprise a series of hydrophobic residues surrounding the cofactor binding site and mutation of both residues negatively affect InhA function. Construction of an M155A mutant of InhA results in increased affinity for NADH and DD-CoA turnover but with a reduction in Vmax for DD-CoA, impairing overall activity. This suggests that NADH-binding geometry of InhA likely permits long-range interactions between residues in the NADH-binding pocket to facilitate substrate turnover in the DD-CoA binding region of the protein. Understanding the precise details of substrate binding and turnover in InhA and how this may affect protein-protein interactions may facilitate the development of improved inhibitors enabling the development of novel anti-TB drugs.
format	article
author	Daniel J. Shaw Kirsty Robb Beatrice V. Vetter Madeline Tong Virginie Molle Neil T. Hunt Paul A. Hoskisson
author_facet	Daniel J. Shaw Kirsty Robb Beatrice V. Vetter Madeline Tong Virginie Molle Neil T. Hunt Paul A. Hoskisson
author_sort	Daniel J. Shaw
title	Disruption of key NADH-binding pocket residues of the Mycobacterium tuberculosis InhA affects DD-CoA binding ability
title_short	Disruption of key NADH-binding pocket residues of the Mycobacterium tuberculosis InhA affects DD-CoA binding ability
title_full	Disruption of key NADH-binding pocket residues of the Mycobacterium tuberculosis InhA affects DD-CoA binding ability
title_fullStr	Disruption of key NADH-binding pocket residues of the Mycobacterium tuberculosis InhA affects DD-CoA binding ability
title_full_unstemmed	Disruption of key NADH-binding pocket residues of the Mycobacterium tuberculosis InhA affects DD-CoA binding ability
title_sort	disruption of key nadh-binding pocket residues of the mycobacterium tuberculosis inha affects dd-coa binding ability
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/969f64fbcfb949d18b2dcc5f36be6f93
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Disruption of key NADH-binding pocket residues of the Mycobacterium tuberculosis InhA affects DD-CoA binding ability

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