Random Mutagenesis of the Prokaryotic Peptide Transporter YdgR Identifies Potential Periplasmic Gating Residues

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2011 May 12

PMID 21558271

Citations 8

Authors

Elisabeth Malle

Hongwen Zhou

Jana Neuhold

Bettina Spitzenberger

Freya Klepsch

Thomas Pollak

Oliver Bergner

Gerhard F Ecker

Peggy C Stolt-Bergner

Affiliations

Soon will be listed here.

Abstract

The peptide transporter (PTR) family represents a group of proton-coupled secondary transporters responsible for bulk uptake of amino acids in the form of di- and tripeptides, an essential process employed across species ranging from bacteria to humans. To identify amino acids critical for peptide transport in a prokaryotic PTR member, we have screened a library of mutants of the Escherichia coli peptide transporter YdgR using a high-throughput substrate uptake assay. We have identified 35 single point mutations that result in a full or partial loss of transport activity. Additional analysis, including homology modeling based on the crystal structure of the Shewanella oneidensis peptide transporter PepT(so), identifies Glu(56) and Arg(305) as potential periplasmic gating residues. In addition to providing new insights into transport by members of the PTR family, these mutants provide valuable tools for further study of the mechanism of peptide transport.

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