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Heme is a cofactor with myriad roles and essential to almost all living organisms. Beyond classical gas transport and catalytic functions, heme is increasingly appreciated as a tightly controlled signalling molecule regulating protein expression. However, heme acquisition, biosynthesis and regulation is poorly understood beyond a few model organisms, and the heme-binding proteome has not been fully characterised in bacteria. Yet as heme homeostasis is critical for bacterial survival, heme-binding proteins are promising drug targets. Herein we report a chemical proteomics method for global profiling of heme-binding proteins in live cells for the first time. Employing a panel of heme-based clickable and photoaffinity probes enabled the profiling of 32-54 % of the known heme-binding proteomes in Gram-positive and Gram-negative bacteria. This simple-to-implement profiling strategy could be interchangeably applied to different cell types and systems and fuel future research into heme biology. © 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.


Isabel V L Wilkinson, Max Bottlinger, Yassmine El Harraoui, Stephan A Sieber. Profiling the Heme-Binding Proteomes of Bacteria Using Chemical Proteomics. Angewandte Chemie (International ed. in English). 2023 Feb 20;62(9):e202212111

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PMID: 36495310

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