A structural characterization of the isoniazidMycobacterium tuberculosisdrug target, Rv2971, in its unliganded form

Abstract

Aldo-keto reductases (AKR) are a large superfamily of NADPH-dependent oxidoreductases and play a role in detoxification of toxic metabolites. Rv2971, an AKR inMycobacterium tuberculosis, has recently been identified as a target of isoniazid, a key first-line drug against tuberculosis. Here, the cloning, expression, purification, crystallization and structural characterization of Rv2971 are described. To gain insight into its function, the crystal structure of Rv2971 was successfully determined to 1.60 Å resolution in its unliganded form. The structure exhibits a TIM-barrel fold typical of AKRs, revealing structural characteristics essential for function and substrate specificities, allowing a structural comparison between Rv2971 and other mycobacterial AKRs.

Using a Label Free Quantitative Proteomics Approach to Identify Changes in Protein Abundance in Multidrug-Resistant Mycobacterium tuberculosis

“…Proteome-wide profiling approach for isoniazid targets in M. tuberculosis has shown that Rv2971 was one of the targets of INH adducts [9]. Recently, the crystal structure of Rv2971 in its unliganded form has been successfully determined and revealed the complex of this protein with the INH-NADH adduct [50]. Therefore, it is possible that the up-regulation of this protein in multidrugresistant strains observed in the present study is necessary for resistance by binding and neutralizing the drug.…”