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

作者：Hammann Philippe; Parmentier Delphine; Cerciat Marie; Reimegard Johan; Helfer Anne Catherine; Boisset Sandrine; Guillier Maude; Vandenesch Francois; Wagner E Gerhart H; Romby Pascale; Fechter Pierre^*

来源：Biochimie, 2014, 106: 175-179.

DOI：10.1016/j.biochi.2014.07.011

• Non-coding RNAs
• DIGE
• Surface proteins
• Post-transcriptional regulation

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 colt, which regulates the synthesis of several outer membrane proteins, and highlighted the role of Staphylococcus aureus RNAIII for the maintenance of cell wall integrity.

• 出版日期2014-11