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

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)
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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 languageEnglish
Article number23191
JournalScientific Reports
Volume6
DOIs
Publication statusPublished - 15 Mar 2016
Externally publishedYes

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.

ASJC Scopus subject areas

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