An Untargeted Metabolomics Strategy to Identify Substrates of Known and Orphan E. coli Transporters.

Radi, Mohammad S, Munro, Lachlan J, Rago, Daniela and Kell, Douglas B ORCID: 0000-0001-5838-7963 (2024) An Untargeted Metabolomics Strategy to Identify Substrates of Known and Orphan E. coli Transporters. Membranes, 14 (3). p. 70.

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Official URL: http://dx.doi.org/10.3390/membranes14030070

Abstract

Transport systems play a pivotal role in bacterial physiology and represent potential targets for medical and biotechnological applications. However, even in well-studied organisms like Escherichia coli, a notable proportion of transporters, exceeding as many as 30%, remain classified as orphans due to their lack of known substrates. This study leveraged high-resolution LC-MS-based untargeted metabolomics to identify candidate substrates for these orphan transporters. Human serum, including a diverse array of biologically relevant molecules, served as an unbiased source for substrate exposure. The analysis encompassed 26 paired transporter mutant contrasts (i.e., knockout vs. overexpression), compared with the wild type, revealing distinct patterns of substrate uptake and excretion across various mutants. The convergence of candidate substrates across mutant scenarios provided robust validation, shedding light on novel transporter-substrate relationships, including those involving yeaV, hsrA, ydjE, and yddA. Furthermore, several substrates were contingent upon the specific mutants employed. This investigation underscores the utility of untargeted metabolomics for substrate identification in the absence of prior knowledge and lays the groundwork for subsequent validation experiments, holding significant implications for both medical and biotechnological advancements.

Item Type:	Article
Uncontrolled Keywords:	E. coli, human serum, orphan transporters, untargeted metabolomics, y-ome
Divisions:	Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology
Depositing User:	Symplectic Admin
Date Deposited:	09 Apr 2024 08:38
Last Modified:	09 Apr 2024 13:16
DOI:	10.3390/membranes14030070
Open Access URL:	https://doi.org/10.3390/membranes14030070
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3180158