2WTZ

MurE ligase of Mycobacterium Tuberculosis

Summary for 2WTZ

• Entry DOI: 10.2210/pdb2wtz/pdb
• Descriptor: UDP-N-ACETYLMURAMOYL-L-ALANYL-D-GLUTAMATE--2,6-DIAMINOPIMELATE LIGASE, URIDINE-5'-DIPHOSPHATE-N-ACETYLMURAMOYL-L-ALANINE-D-GLUTAMATE, MAGNESIUM ION (3 entities in total)
• Functional Keywords: nucleotide-binding, peptidoglycan synthesis, mure, ligase, cell shape, cell cycle, mycobacterium tuberculosis, cell wall biogenesis/degradation, atp-binding, peptidoglycan, cell division
• Biological source: MYCOBACTERIUM TUBERCULOSIS
• Cellular location: Cytoplasm (By similarity): P65477
• Total number of polymer chains: 4
• Total formula weight: 225434.15

Authors

Basavannacharya, C.,Robertson, G.,Munshi, T.,Keep, N.H.,Bhakta, S. (deposition date: 2009-09-25, release date: 2009-12-15, Last modification date: 2023-12-20)

Primary citation

Basavannacharya, C.,Robertson, G.,Munshi, T.,Keep, N.H.,Bhakta, S.
ATP-Dependent Mure Ligase in Mycobacterium Tuberculosis: Biochemical and Structural Characterisation.
Tuberculosis(Edinb.), 90:16-, 2010

PubMed Abstract: New therapies are required against Mycobacterium tuberculosis and its cell wall peptidoglycan biosynthesis is a potential therapeutic target. UDP-MurNAc-tripeptide ligase (MurE) is a member of the ATP-dependent ligase family, which incorporate amino acids including meso-diaminopimelic acid (m-DAP) into peptidoglycan during synthesis in a species-specific manner. In the present study, we have cloned, over-expressed, and characterised MurE from M. tuberculosis (Mtb-MurE). The crystal structure has been determined at 3.0A resolution in the presence of the substrate UDP-MurNAc-l-Ala-d-Glu (UAG). The activity of the enzyme was measured through estimating inorganic phosphate released in a non-radioactive high-throughput colourimetric assay. UDP-MurNAc-l-Ala-d-Glu-m-DAP (UMT) formation coupled to inorganic phosphate release was confirmed by HPLC and mass spectrometric analyses. Kinetic constants were determined for a range of natural substrates using optimised conditions. From our findings, it is evident that Mtb-MurE is highly specific in adding m-DAP to UDP-MurNAc-dipeptide and ATP-hydrolysis is an absolute requirement for its activity.

PubMed: 19945347
DOI: 10.1016/J.TUBE.2009.10.007

Experimental method

X-RAY DIFFRACTION (3 Å)