Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/99891
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Type: Journal article
Title: Isolation and identification of Enterococcus faecalis membrane proteins using membrane shaving, 1D SDS/PAGE, and mass spectrometry
Author: Cathro, P.
Mccarthy, P.
Hoffmann, P.
Zilm, P.
Citation: FEBS Open Bio, 2016; 6(6):586-593
Publisher: Febs Press
Issue Date: 2016
ISSN: 2211-5463
2211-5463
Statement of
Responsibility: 
Peter Cathro, Peter McCarthy, Peter Hoffmann, and Peter Zilm
Abstract: Enterococcus faecalis is a significant nosocomial pathogen, which is able to survive in diverse environments and resist killing with antimicrobial therapies. The expression of cell membrane proteins play an important role in how bacteria respond to environmental stress. As such, the capacity to identify and study membrane protein expression is critical to our understanding of how specific proteins influence bacterial survival. Here, we describe a combined approach to identify membrane proteins of E. faecalis ATCC V583 using membranes fractionated by either 1D SDS/PAGE or membrane shaving, coupled with LC-ESI mass spectrometry. We identified 222 membrane-associated proteins, which represent approximately 24% of the predicted membrane-associated proteome: 170 were isolated using 1D SDS/PAGE and 68 with membrane shaving, with 36 proteins being common to both the techniques. Of the proteins identified by membrane shaving, 97% were membrane-associated with the majority being integral membrane proteins (89%). Most of the proteins identified with known physiology are involved with transportation across the membrane. The combined 1D SDS/PAGE and membrane shaving approach has produced the greatest number of membrane proteins identified from E. faecalis to date. These protocols will aid future researchers investigating changes in the membrane proteome of E. faecalis by improving our understanding of how E. faecalis adapts and responds to its environment.
Keywords: 1D SDS/PAGE; Enterococcus faecalis; mass spectrometry; membrane shaving; proteomics
Rights: © 2016 The Authors. Published by FEBS Press and John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
RMID: 0030049493
DOI: 10.1002/2211-5463.12075
Appears in Collections:Molecular and Biomedical Science publications

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