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: 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
• Journal: Scientific reports (Sci Rep) Vol. 6 Pg. 23191 (Mar 15 2016) ISSN: 2045-2322 [Electronic] England
• PMID: 26976706 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Bacterial Proteins
• His-His-His-His-His-His
• Oligopeptides
• Histidine
• Diaminopimelic Acid
• Captopril
• Amidohydrolases
• Zinc

Topics

• Amidohydrolases (antagonists & inhibitors, chemistry, metabolism)
• Bacterial Proteins (chemistry, metabolism)
• Binding Sites
• Captopril (pharmacology)
• Diaminopimelic Acid (chemistry, metabolism)
• Histidine (chemistry)
• Mycobacterium tuberculosis (enzymology)
• Oligopeptides (chemistry)
• Zinc (chemistry)