Reconstruction of diaminopimelic acid biosynthesis allows characterisation of Mycobacterium tuberculosis N-succinyl-L,L-diaminopimelic acid desuccinylase

Veeraraghavan Usha; Adrian J. Lloyd; David I. Roper; Christopher G. Dowson; Guennadi Kozlov; Kalle Gehring; Smita Chauhan; Hasan T. Imam; Claudia A. Blindauer; Gurdyal S. Besra

doi:10.1038/srep23191

Reconstruction of diaminopimelic acid biosynthesis allows characterisation of Mycobacterium tuberculosis N-succinyl-L,L-diaminopimelic acid desuccinylase

Veeraraghavan Usha, Adrian J. Lloyd^*, David I. Roper, Christopher G. Dowson, Guennadi Kozlov, Kalle Gehring, Smita Chauhan, Hasan T. Imam, Claudia A. Blindauer, Gurdyal S. Besra

^*Corresponding author for this work

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Abstract

With the increased incidence of tuberculosis (TB) caused by Mycobacterium tuberculosis there is an urgent need for new and better anti-tubercular drugs. N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE) is a key enzyme in the succinylase pathway for the biosynthesis of meso-diaminopimelic acid (meso-DAP) and L-lysine. DapE is a zinc containing metallohydrolase which hydrolyses N-succinyl L,L diaminopimelic acid (L,L-NSDAP) to L,L-diaminopimelic acid (L,L-DAP) and succinate. M. tuberculosis DapE (MtDapE) was cloned, over-expressed and purified as an N-terminal hexahistidine ((His) 6) tagged fusion containing one zinc ion per DapE monomer. We redesigned the DAP synthetic pathway to generate L,L-NSDAP and other L,L-NSDAP derivatives and have characterised MtDapE with these substrates. In contrast to its other Gram negative homologues, the MtDapE was insensitive to inhibition by L-captopril which we show is consistent with novel mycobacterial alterations in the binding site of this drug.

Original language	English
Article number	23191
Journal	Scientific Reports
Volume	6
DOIs	https://doi.org/10.1038/srep23191
Publication status	Published - 15 Mar 2016
Externally published	Yes

Bibliographical note

Funding Information:
The authors gratefully acknowledge the support of the Wellcome Trust, MRC and the Birmingham-Warwick Science City Translational Medicine Initiative. We would also like to gratefully acknowledge the assistance of Dr. Susan Slade of the Warwick/Waters Centre for BioMedical Mass Spectrometry and Proteomics in the School of Life Sciences, University of Warwick. AJL was supported by a Birmingham Science City Interdisciplinary Research Alliance Fellowship.

Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.

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10.1038/srep23191Licence: CC BY

Reconstruction of diaminopimelic acid biosynthesis allows characterisation of Mycobacterium tuberculosis N-succinyl-L,Ldiaminopimelic acid desuccinylase
© 2021 The Authors. This is an open access article published under a Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
Final published version, 1.98 MBLicence: CC BY

Cite this

Usha, V., Lloyd, A. J., Roper, D. I., Dowson, C. G., Kozlov, G., Gehring, K., Chauhan, S., Imam, H. T., Blindauer, C. A., & Besra, G. S. (2016). Reconstruction of diaminopimelic acid biosynthesis allows characterisation of Mycobacterium tuberculosis N-succinyl-L,L-diaminopimelic acid desuccinylase. Scientific Reports, 6, Article 23191. https://doi.org/10.1038/srep23191

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title = "Reconstruction of diaminopimelic acid biosynthesis allows characterisation of Mycobacterium tuberculosis N-succinyl-L,L-diaminopimelic acid desuccinylase",

abstract = "With the increased incidence of tuberculosis (TB) caused by Mycobacterium tuberculosis there is an urgent need for new and better anti-tubercular drugs. N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE) is a key enzyme in the succinylase pathway for the biosynthesis of meso-diaminopimelic acid (meso-DAP) and L-lysine. DapE is a zinc containing metallohydrolase which hydrolyses N-succinyl L,L diaminopimelic acid (L,L-NSDAP) to L,L-diaminopimelic acid (L,L-DAP) and succinate. M. tuberculosis DapE (MtDapE) was cloned, over-expressed and purified as an N-terminal hexahistidine ((His) 6) tagged fusion containing one zinc ion per DapE monomer. We redesigned the DAP synthetic pathway to generate L,L-NSDAP and other L,L-NSDAP derivatives and have characterised MtDapE with these substrates. In contrast to its other Gram negative homologues, the MtDapE was insensitive to inhibition by L-captopril which we show is consistent with novel mycobacterial alterations in the binding site of this drug.",

author = "Veeraraghavan Usha and Lloyd, {Adrian J.} and Roper, {David I.} and Dowson, {Christopher G.} and Guennadi Kozlov and Kalle Gehring and Smita Chauhan and Imam, {Hasan T.} and Blindauer, {Claudia A.} and Besra, {Gurdyal S.}",

note = "Funding Information: The authors gratefully acknowledge the support of the Wellcome Trust, MRC and the Birmingham-Warwick Science City Translational Medicine Initiative. We would also like to gratefully acknowledge the assistance of Dr. Susan Slade of the Warwick/Waters Centre for BioMedical Mass Spectrometry and Proteomics in the School of Life Sciences, University of Warwick. AJL was supported by a Birmingham Science City Interdisciplinary Research Alliance Fellowship. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

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AU - Dowson, Christopher G.

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AU - Blindauer, Claudia A.

AU - Besra, Gurdyal S.

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Reconstruction of diaminopimelic acid biosynthesis allows characterisation of Mycobacterium tuberculosis N-succinyl-L,L-diaminopimelic acid desuccinylase

Abstract

Bibliographical note

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UN SDGs

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