PMID:15812637
| Citation |
Norman, C, Liu, ZW, Rigby, P, Raso, A, Petrov, Y and Martinac, B (2005) Visualisation of the mechanosensitive channel of large conductance in bacteria using confocal microscopy. Eur. Biophys. J. 34:396-402 |
|---|---|
| Abstract |
The mechanosensitive channel of large conductance (MscL) plays an important role in the survival of bacterial cells to hypo-osmotic shock. This channel has been extensively studied and its sequence, structure and electrophysiological characteristics are well known. Here we present a method to visualise MscL in living bacteria using confocal microscopy. By creating a gene fusion between mscl and the gene encoding the green fluorescent protein (GFP) we were able to express the fusion protein MscL-GFP in bacteria. We show that MscL-GFP is present in the cytoplasmic membrane and forms functional channels. These channels have the same characteristics as wild-type MscL, except that they require more pressure to open. This method could prove an interesting, non-invasive, tool to study the localisation and the regulation of expression of MscL in bacteria. |
| Links |
PubMed Online version:10.1007/s00249-005-0475-y |
| Keywords |
Bacterial Physiological Phenomena; Bacterial Proteins/chemistry; Biophysics/instrumentation; Biophysics/methods; Cell Membrane/metabolism; Cell Proliferation; Cytoplasm/metabolism; Electrophysiology; Escherichia coli/metabolism; Escherichia coli Proteins/chemistry; Green Fluorescent Proteins/chemistry; Green Fluorescent Proteins/metabolism; Ion Channels/chemistry; Microscopy, Confocal/methods; Plasmids/metabolism; Pressure; Recombinant Fusion Proteins/chemistry; Spheroplasts/metabolism; Time Factors |
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