Interaction of the transmembrane domain of lysis protein E from bacteriophage phiX174 with bacterial translocase MraY and peptidyl-prolyl isomerase SlyD

Sharon Mendel, Joanne M Holbourn, James A Schouten, Timothy D H Bugg

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29 Citations (Scopus)

Abstract

The molecular target for the bacteriolytic E protein from bacteriophage X174, responsible for host cell lysis, is known to be the enzyme phospho-MurNAc-pentapeptide translocase (MraY), an integral membrane protein involved in bacterial cell wall peptidoglycan biosynthesis, with an essential role being played by peptidyl-prolyl isomerase SlyD. A synthetic 37 aa peptide E(pep), containing the N-terminal transmembrane alpha-helix of E, was found to be bacteriolytic against Bacillus licheniformis, and inhibited membrane-bound MraY. The solution conformation of E(pep) was found by circular dichroism (CD) spectroscopy to be 100 % alpha-helical. No change in the CD spectrum was observed upon addition of purified Escherichia coli SlyD, implying that SlyD does not catalyse prolyl isomerization upon E. However, E(pep) was found to be a potent inhibitor of SlyD-catalysed peptidyl-prolyl isomerization (IC(50) 0.15 microM), implying a strong interaction between E and SlyD. E(pep) was found to inhibit E. coli MraY activity when assayed in membranes (IC(50) 0.8 microM); however, no inhibition of solubilized MraY was observed, unlike nucleoside natural product inhibitor tunicamycin. These results imply that the interaction of E with MraY is not at the MraY active site, and suggest that a protein-protein interaction is formed between E and MraY at a site within the transmembrane region.

Original languageEnglish
Pages (from-to)2959-2967
Number of pages9
Journal BMC Microbiology
Volume152
Issue numberPt 10
Early online date1 Oct 2006
DOIs
Publication statusPublished - Oct 2006

Keywords

  • Anti-Bacterial Agents
  • Bacillus
  • Bacterial Proteins
  • Bacteriolysis
  • Circular Dichroism
  • Escherichia coli Proteins
  • Models, Biological
  • Peptides
  • Peptidylprolyl Isomerase
  • Protein Binding
  • Protein Interaction Mapping
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Transferases
  • Viral Proteins
  • Journal Article
  • Research Support, Non-U.S. Gov't

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