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New insights into the substrate specificities of proton-coupled oligopeptide transporters from E. coli by a pH sensitive assay
Authors:Bala K. Prabhala  Nanda G. Aduri  Johanne M. Jensen  Heidi A. Ernst  Nida Iram  Moazur Rahman  Osman Mirza
Affiliation:1. Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark;2. Health Biotechnology Divisions, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan
Abstract:Proton-coupled oligopeptide transporters (POTs) are secondary active transporters that facilitate di- and tripeptide uptake by coupling it to an inward directed proton electrochemical gradient. Here the substrate specificities of Escherichia coli POTs YdgR, YhiP and YjdL were investigated by means of a label free transport assay using the hydrophilic pH sensitive dye pyranine and POT overexpressing E. coli cells. The results confirm and extend the functional knowledge on E. coli POTs. In contrast to previous assumptions, alanine and trialanine appears to be substrates of YjdL, albeit poor compared to dipeptides. Similarly tetraalanine apparently is a substrate of both YdgR and YhiP.
Keywords:Pyranine   POTs   Peptides   Proton   Bioassay
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