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.
authors.affiliation: School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
authors.forename: Veeraraghavan
authors.initials: V
authors.lastname: Usha
authors.affiliation: School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK.
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Lloyd
authors.affiliation: School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK.
authors.forename: David I
authors.initials: DI
authors.lastname: Roper
authors.affiliation: School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Dowson
authors.affiliation: Department of Biochemistry, McGill University, Montreal, Quebec, H3G 1Y6, Canada.
authors.forename: Guennadi
authors.initials: G
authors.lastname: Kozlov
authors.affiliation: Department of Biochemistry, McGill University, Montreal, Quebec, H3G 1Y6, Canada.
authors.forename: Kalle
authors.initials: K
authors.lastname: Gehring
authors.affiliation: School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK.
authors.forename: Smita
authors.initials: S
authors.lastname: Chauhan
authors.affiliation: Department of Chemistry, University of Warwick, Coventry, CV4 7AL,UK.
authors.forename: Hasan T
authors.initials: HT
authors.lastname: Imam
authors.affiliation: Department of Chemistry, University of Warwick, Coventry, CV4 7AL,UK.
authors.forename: Claudia A
authors.initials: CA
authors.lastname: Blindauer
authors.affiliation: School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
authors.forename: Gurdyal S
authors.initials: GS
authors.lastname: Besra
mesh_terms: D000581:Amidohydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D002216:Captopril / Q000494:pharmacology; D003960:Diaminopimelic Acid / Q000737:chemistry / Q000378:metabolism*; D006639:Histidine / Q000737:chemistry; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D009842:Oligopeptides / Q000737:chemistry; D015032:Zinc / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; C471213:His-His-His-His-His-His; D009842:Oligopeptides; D006639:Histidine; D003960:Diaminopimelic Acid; D002216:Captopril; D000581:Amidohydrolases; D015032:Zinc