Visualisation of the mechanosensitive channel of large conductance in bacteria using confocal microscopy

Norman, Christel, Liu, Zhen-Wei, Rigby, Paul, Raso, Albert, Petrov, Yevgeniy and Martinac, Boris (2005) Visualisation of the mechanosensitive channel of large conductance in bacteria using confocal microscopy. European Biophysics Journal, 34 5: 396-402. doi:10.1007/s00249-005-0475-y


Author Norman, Christel
Liu, Zhen-Wei
Rigby, Paul
Raso, Albert
Petrov, Yevgeniy
Martinac, Boris
Title Visualisation of the mechanosensitive channel of large conductance in bacteria using confocal microscopy
Journal name European Biophysics Journal   Check publisher's open access policy
ISSN 0175-7571
1432-1017
Publication date 2005-01-01
Sub-type Article (original research)
DOI 10.1007/s00249-005-0475-y
Volume 34
Issue 5
Start page 396
End page 402
Total pages 7
Editor A. Watts
Place of publication Berlin, Germany
Publisher Springer
Language eng
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.
Keyword Mechanosensitive channel of large conductance
Green fluorescent protein
Confocal microscopy
Patch-clamp
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ
Additional Notes Published online: 6 April 2005

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
School of Biomedical Sciences Publications
 
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Created: Thu, 20 Sep 2007, 04:15:01 EST