Here, we report the synthesis of new CyC alkyne-containing inhibitors (i.e., CyCyne) and their use for the direct capture of their target proteins in M. tuberculosis cultures via a bio-orthogonal click-chemistry activity-based protein profiling (CC-ABPP) approach combined with mass spectrometry.

This strategy led to the identification of around 160 potential target enzymes, most of them involved in lipid metabolism and/or in cell wall biosynthesis. One of these enzymes, HsaD (Rv3569c), is required for M. tb survival within macrophages. The specificity of HsaD inhibition by various CyC analogues was deciphered and validated by X-ray structures of four HsaD-CyC complexes in the 1.6-2.6 Å resolution range.

Sarah Barelier, Romain Avellan, Giri Raj Gnawali, Patrick Fourquet, Véronique Roig-Zamboni, Isabelle Poncin, Vanessa Point, Yves Bourne, Stéphane Audebert, Luc Camoin, Christopher Spilling, Stéphane Canaan, Jean‐françois Cavalier, Gerlind Sulzenbacher

FEBS Journal (2022), DOI: 10.1111/febs.16645

Published on 17/11/2022

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