Bioassay-Driven, Fractionation-Empowered, Focused Metabolomics for Discovering Bacterial Activators of Aryl Hydrocarbon Receptor.
Document Type
Article
Publication Date
3-6-2024
Original Citation
Tian H,
Wang L,
Hardy R,
Kozhaya L,
Placek L,
Fleming E,
Oh J,
Unutmaz D,
Yao X.
Bioassay-Driven, Fractionation-Empowered, Focused Metabolomics for Discovering Bacterial Activators of Aryl Hydrocarbon Receptor. J Am Soc Mass Spectrom. 2024 Mar 6;35(3):518-526.
Keywords
JGM, Humans, Receptors, Aryl Hydrocarbon, Tandem Mass Spectrometry
JAX Source
J Am Soc Mass Spectrom. 2024 Mar 6;35(3):518-526.
ISSN
1879-1123
PMID
38308645
DOI
https://doi.org/10.1021/jasms.3c00386
Abstract
Aryl hydrocarbon receptor (AhR) is a transcription factor that regulates gene expression upon ligand activation, enabling microbiota-dependent induction, training, and function of the host immune system. A spectrum of metabolites, encompassing indole and tryptophan derivatives, have been recognized as activators. This work introduces an integrated, mass spectrometry-centric workflow that employs a bioassay-guided, fractionation-based methodology for the identification of AhR activators derived from human bacterial isolates. By leveraging the workflow efficiency, the complexities inherent in metabolomics profiling are significantly reduced, paving the way for an in-depth and focused mass spectrometry analysis of bioactive fractions isolated from bacterial culture supernatants. Validation of AhR activator candidates used multiple criteria─MS/MS of the synthetic reference compound, bioassay of AhR activity, and elution time confirmation using a C-13 isotopic reference─and was demonstrated for N-formylkynurenine (NFK). The workflow reported provides a roadmap update for improved efficiency of identifying bioactive metabolites using mass spectrometry-based metabolomics. Mass spectrometry datasets are accessible at the National Metabolomics Data Repository (PR001479, Project DOI: 10.21228/M8JM7Q).