Biological and Medicinal Chemistry

Eukaryotic catecholamine hormones influence the chemotactic control of Vibrio campbellii by binding to the coupling protein CheW

Authors

Abstract

In addition to their well-known role as stress-associated catecholamine hormones in animals and humans, epinephrine (EPI) and norepinephrine (NE) act as interkingdom signals between eukaryotic hosts and bacteria. However, the molecular basis of their effects on bacteria is not well understood. In initial phenotypic studies utilizing Vibrio campbellii as a model organism, we characterized the bipartite mode of action of catecholamines, which consists of promotion of growth under iron limitation, and enhanced colony expansion on soft agar. In order to identify the molecular targets of the hormones, we designed and synthesized tailored probes for chemical proteomic studies. As the catechol group in EPI and NE acts as iron chelator and is prone to form a reactive quinone moiety, we devised a photoprobe based on the adrenergic agonist phenylephrine (PE), which solely influenced colony expansion. Using this probe, we identified CheW, located at the core of the chemotaxis signaling network, as a major target. In vitro studies confirmed that EPI, NE, PE, as well as labetalol, a clinically applied antagonist, bind to purified CheW with affinity constants in the sub-micromolar range. In line with these findings, exposure of V. campbellii to these adrenergic agonists affects the chemotactic control of the bacterium. This study highlights a previously unknown effect of eukaryotic signaling molecules on bacterial motility.

Content

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Supplementary material

Thumbnail image of Supplements 15102021 ChemRxiv.pdf
Supplementary Material for Eukaryotic catecholamine hormones influence the chemotactic control of Vibrio campbellii by binding to the coupling protein CheW
This file contains supplementary methods, figures, tables, and legens for supplementary datasets.
Thumbnail image of Table S1 EPI-P1 vs DMSO final.xlsx
Dataset S1
Proteomics data of EPI-P1 photolabeling containing detailed protein annotation, MS data, statistics, and LFQ intensities.
Thumbnail image of Table S2 PE-P vs PE final.xlsx
Dataset S2
Proteomics data of PE-P photolabeling in competition with PE containing detailed protein annotation, MS data, statistics, and LFQ intensities.
Thumbnail image of Table S3 PE-P vs EPI final.xlsx
Dataset S3
Proteomics data of PE-P photolabeling in competition with EPI containing detailed protein annotation, MS data, statistics, and LFQ intensities.
Thumbnail image of Table S4 PE-P vs LAB PRO final.xlsx
Dataset S4
Proteomics data of PE-P photolabeling in competition with LAB or PRO containing detailed protein annotation, MS data, statistics, and LFQ intensities.
Thumbnail image of Table S5 isoDTB final_SMH.xlsx
Dataset S5
Proteomics data of isoDTB labeling experiment containing amino acid modification, selectivity, and quantification results.
Thumbnail image of Table S6 Co-IP 3G final.xlsx
Dataset S6
Proteomics data of Co-IP containing detailed protein annotation, MS data, statistics, and LFQ intensities.