4RMF

Biochemical and structural characterization of mycobacterial aspartyl-tRNA synthetase AspS, a promising TB drug target

4RMF の概要

• エントリーDOI: 10.2210/pdb4rmf/pdb
• 分子名称: Aspartate--tRNA(Asp/Asn) ligase, 2,2-bis(hydroxymethyl)propane-1,3-diol, FORMIC ACID, ... (4 entities in total)
• 機能のキーワード: alpha and beta proteins, trna synthetase, trna, ligase
• 由来する生物種: Mycobacterium smegmatis
• 細胞内の位置: Cytoplasm : A0QWN3
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 66711.91

構造登録者

Cox, J.A.G.,Gurcha, S.S.,Veeraraghavan, U.,Besra, G.S.,Futterer, K. (登録日: 2014-10-21, 公開日: 2014-11-19, 最終更新日: 2023-09-20)

主引用文献

Gurcha, S.S.,Usha, V.,Cox, J.A.,Futterer, K.,Abrahams, K.A.,Bhatt, A.,Alderwick, L.J.,Reynolds, R.C.,Loman, N.J.,Nataraj, V.,Alemparte, C.,Barros, D.,Lloyd, A.J.,Ballell, L.,Hobrath, J.V.,Besra, G.S.
Biochemical and Structural Characterization of Mycobacterial Aspartyl-tRNA Synthetase AspS, a Promising TB Drug Target.
Plos One, 9:e113568-e113568, 2014

PubMed Abstract: The human pathogen Mycobacterium tuberculosis is the causative agent of pulmonary tuberculosis (TB), a disease with high worldwide mortality rates. Current treatment programs are under significant threat from multi-drug and extensively-drug resistant strains of M. tuberculosis, and it is essential to identify new inhibitors and their targets. We generated spontaneous resistant mutants in Mycobacterium bovis BCG in the presence of 10× the minimum inhibitory concentration (MIC) of compound 1, a previously identified potent inhibitor of mycobacterial growth in culture. Whole genome sequencing of two resistant mutants revealed in one case a single nucleotide polymorphism in the gene aspS at (535)GAC>(535)AAC (D179N), while in the second mutant a single nucleotide polymorphism was identified upstream of the aspS promoter region. We probed whole cell target engagement by overexpressing either M. bovis BCG aspS or Mycobacterium smegmatis aspS, which resulted in a ten-fold and greater than ten-fold increase, respectively, of the MIC against compound 1. To analyse the impact of inhibitor 1 on M. tuberculosis AspS (Mt-AspS) activity we over-expressed, purified and characterised the kinetics of this enzyme using a robust tRNA-independent assay adapted to a high-throughput screening format. Finally, to aid hit-to-lead optimization, the crystal structure of apo M. smegmatis AspS was determined to a resolution of 2.4 Å.

PubMed: 25409504
DOI: 10.1371/journal.pone.0113568

実験手法

X-RAY DIFFRACTION (2.4 Å)