Ravindran, Madhu Sudhan, Rao, Srinivasa P.S., Cheng, Xiamin, Shukla, Ankit, Cazenave-Gassiot, Amaury, Yao, Shao Q. and Wenk, Markus R. (2014) Targeting lipid esterases in mycobacteria grown under different physiological conditions using activity based profiling with tetrahydrolipstatin. Molecular & cellular proteomics : MCP, 13 (2). pp. 435-448. ISSN 1535-9484

Abstract

Tetrahydrolipstatin (THL) is bactericidal but its precise target spectrum is poorly characterized. Here, we used a THL analogue and activity-based protein profiling for identification of target proteins after enrichment from whole cell lysates of Mycobacterium bovis Bacillus Calmette-Guérin (BCG) cultured under growing and non-replicating conditions. THL targets /-hydrolases including many lipid esterases (LipD, G, H, I, M, N, O, V, W, and TesA). Target protein concentrations and total esterase activity correlated inversely with cellular triacylglycerol (TAG) upon entry into and exit from non-replicative conditions. Cellular overexpression of lipH and tesA led to decreased THL susceptibility thus providing functional validation. Our results define the target spectrum of THL in a biological species which has particularly diverse lipid metabolic pathways. We furthermore derive a conceptual approach that demonstrates the utility of such THL probes for the characterization of substrate recognition by lipases and related enzymes.

Item Type:	Article
Keywords:	Activity-Based Protein Profiling, Tetrahydrolipstatin, Mycobacteria, Lipases, Esterases, Mass spectrometry, Lipidomics, Triacylglycerol
Date Deposited:	12 Oct 2016 00:45
Last Modified:	12 Oct 2016 00:45
URI:	https://oak.novartis.com/id/eprint/9681