The prototypical proton-coupled oligopeptide transporter YdgR from Escherichia coli facilitates chloramphenicol uptake into bacterial cells

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Bala K Prabhala, Nanda G Aduri, Neha Sharma, Aqsa Shaheen, Arpan Sharma, Mazhar Iqbal, Paul R Hansen, Christoffer Brasen, Michael Gajhede, Moazur Rahman, Osman Mirza

Chloramphenicol (Cam) is a broad-spectrum antibiotic used to combat bacterial infections in humans and animals. Cam export from bacterial cells is one of the mechanisms by which pathogens resist Cam's antibacterial effects, and several different proteins are known to facilitate this process. However, to date no report exists on any specific transport protein that facilitates Cam uptake. The proton-coupled oligopeptide transporter (POT) YdgR from Escherichia coli is a prototypical member of the POT family, functioning in proton-coupled uptake of di- and tripeptides. By following bacterial growth and conducting LC-MS-based assays we show here that YdgR facilitates Cam uptake. Some YdgR variants displaying reduced peptide uptake also exhibited reduced Cam uptake, indicating that peptides and Cam bind YdgR at similar regions. Homology modeling of YdgR, Cam docking, and mutational studies suggested a binding mode that resembles that of Cam binding to the multidrug resistance transporter MdfA. To our knowledge, this is the first report of Cam uptake into bacterial cells mediated by a specific transporter protein. Our findings suggest a specific bacterial transporter for drug uptake that might be targeted to promote greater antibiotic influx in order to increase cytoplasmic antibiotic concentration for enhanced cytotoxicity.

Original languageEnglish
JournalThe Journal of Biological Chemistry
Volume293
Pages (from-to)1007-1017
ISSN0021-9258
DOIs
Publication statusPublished - 2018

    Research areas

  • Journal Article

ID: 185842508