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Isotopically Labeled Desthiobiotin Azide (isoDTB) Tags Enable Global Profiling of the Bacterial Cysteinome

preprint
submitted on 16.09.2019 and posted on 20.09.2019 by Patrick R. A. Zanon, Lisa Lewald, Stephan M. Hacker
Rapid development of bacterial resistance has led to an urgent need to find new druggable targets for antibiotics. In this context, residue-specific chemoproteomic approaches enable proteome-wide identification of binding sites for covalent inhibitors. Here, we describe isotopically labeled desthiobiotin azide (isoDTB) tags that are easily synthesized, shorten the chemoproteomic workflow and allow an increased coverage of cysteines in bacterial systems. We quantify 59% of all cysteines in essential proteins in Staphylococcus aureus and discover 88 cysteines with high reactivity, which correlates with functional importance. Furthermore, we identify 268 cysteines that are engaged by covalent ligands. We verify inhibition of HMG-CoA synthase, which will allow addressing the bacterial mevalonate pathway through a new target. Overall, a comprehensive map of the bacterial cysteinome is obtained, which will facilitate the development of antibiotics with novel modes-of-action.

Funding

Liebig Fellowship by the Fonds der Chemischen Industrie

Ph.D. fellowship by the Fonds der Chemischen Industrie

TUM Junior Fellow Fund

History

Email Address of Submitting Author

stephan.m.hacker@tum.de

Institution

Technical University of Munich

Country

Germany

ORCID For Submitting Author

0000-0001-5420-4824

Declaration of Conflict of Interest

The authors declare no conflict of interest.

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