A method to map changes in bacterial surface composition induced by regulatory RNAs in Escherichia coli and Staphylococcus aureus

Philippe Hammann, Delphine Parmentier, Marie Cerciat, Johan Reimegård, Anne-Catherine Helfer, Sandrine Boisset, Maude Guillier, François Vandenesch, E. Gerhart H. Wagner, Pascale Romby, Pierre Fechter
2014 Biochimie  
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
more » ... scherichia 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.
doi:10.1016/j.biochi.2014.07.011 pmid:25046628 fatcat:tdnw5l27gzc7led25ksddy625a