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Volume 152, Issue 10

Interaction of the transmembrane domain of lysis protein E from bacteriophage X174 with bacterial translocase MraY and peptidyl-prolyl isomerase SlyD Free

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, containing the N-terminal transmembrane -helix of E, was found to be bacteriolytic against , and inhibited membrane-bound MraY. The solution conformation of E was found by circular dichroism (CD) spectroscopy to be 100 % -helical. No change in the CD spectrum was observed upon addition of purified SlyD, implying that SlyD does not catalyse prolyl isomerization upon E. However, E was found to be a potent inhibitor of SlyD-catalysed peptidyl-prolyl isomerization (IC 0.15 μM), implying a strong interaction between E and SlyD. E was found to inhibit MraY activity when assayed in membranes (IC 0.8 μM); 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.

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Keyword(s): CD, circular dichroism , FKBP, FK506-binding protein and MraY, phospho-MurNAc-pentapeptide translocase
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2006-10-01
2024-04-19
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Interaction of the transmembrane domain of lysis protein E from bacteriophage ϕX174 with bacterial translocase MraY and peptidyl-prolyl isomerase SlyD
Microbiology 152, 2959 (2006); https://doi.org/10.1099/mic.0.28776-0
/content/journal/micro/10.1099/mic.0.28776-0
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