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A method to map changes in bacterial surface composition induced by regulatory RNAs in Escherichia coli and Staphylococcus aureus
Ist Teil von
Biochimie, 2014-11, Vol.106, p.175-179
Ort / Verlag
France: Elsevier B.V
Erscheinungsjahr
2014
Link zum Volltext
Quelle
Elsevier ScienceDirect Journals Complete
Beschreibungen/Notizen
We have adapted a method to map cell surface proteins and to monitor the effect of specific regulatory RNAs on the surface composition of the bacteria. This method involves direct labeling of surface proteins of living bacteria using fluorescent dyes and a subsequent separation of the crude extract by 2D gel electrophoresis. The strategy yields a substantial enrichment in surface proteins over cytoplasmic proteins. We validated this method by monitoring the effect of the regulatory RNA MicA in Escherichia coli, which regulates the synthesis of several outer membrane proteins, and highlighted the role of Staphylococcus aureus RNAIII for the maintenance of cell wall integrity.
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•Surface proteins of live cells were labeled with Cy-Dyes and identified by mass spectrometry.•The strategy was validated to monitor the effect of regulatory RNAs on the surface composition of bacteria.•Novel targets for Escherichia coli MicA were highlighted.•Staphylococcus aureus RNAIII is required to maintain the cell wall integrity.