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Updated: Apr 26, 2026

MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria
Published on: February 23, 2021
A method to map changes in bacterial surface composition induced by regulatory RNAs in Escherichia coli and
Philippe Hammann1, Delphine Parmentier2, Marie Cerciat2
1Esplanade Proteomic Platform, Université de Strasbourg, CNRS, IBMC, 15 Rue René Descartes, F-67084 Strasbourg, France.
Abstract:
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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