Physiological response in E. coli to YdgR overexpression depends on whether the protein has an intact function

Salvia Sajid, Lilia Hernandez Salas, Maria Rafiq, Torben Lund, Mikkel Girke Jørgensen, Bent Honoré, Lars Porskjær Christensen, Paul Robert Hansen, Henrik Franzyk, Osman Mirza, Bala Krishna Prabhala

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Membrane transport proteins are essential for the transport of a wide variety of molecules across the cell membrane to maintain cellular homeostasis. Generally, these transport proteins can be overexpressed in a suitable host (bacteria, yeast, or mammalian cells), and it is well documented that overexpression of membrane proteins alters the global metabolomic and proteomic profiles of the host cells. In the present study, we investigated the physiological consequences of overexpression of a membrane transport protein YdgR that belongs to the POT/PTR family from E. coli by using the lab strain BL21 (DE3)pLysS in its functional and attenuated mutant YdgR-E33Q. We found significant differences between the omics (metabolomics and proteomics) profiles of the cells expressing functional YdgR as compared to cells expressing attenuated YdgR, e.g., upregulation of several uncharacterized y-proteins and enzymes involved in the metabolism of peptides and amino acids. Furthermore, molecular network analysis suggested a relatively higher presence of proline-containing tripeptides in cells expressing functional YdgR. We envisage that an in-depth investigation of physiological alterations due to protein over-expression may be used for the deorphanization of the y-gene transportome.
Original languageEnglish
JournalBiochemical and Biophysical Research Communications
Pages (from-to)42-49
Number of pages8
Publication statusPublished - 18. Jun 2023


  • E. coli BL21(DE3)pLysS
  • Metabolomics/proteomics
  • POT/PTR family
  • Transport protein overexpression
  • YdgR
  • Membrane Transport Proteins/metabolism
  • Escherichia coli/metabolism
  • Recombinant Proteins/metabolism
  • Animals
  • Proteomics
  • Mammals/metabolism
  • Escherichia coli Proteins/metabolism
  • Carrier Proteins/metabolism


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