Visualisation of the mechanosensitive channel of large conductance in bacteria using confocal microscopy |
| |
Authors: | Christel Norman Zhen-Wei Liu Paul Rigby Albert Raso Yevgeniy Petrov Boris Martinac |
| |
Affiliation: | (1) Pharmacology Unit M510, School of Medicine and Pharmacology, QEII Medical Centre, The University of Western Australia, Crawley, WA, 6009, Australia;(2) Biomedical Imaging and Analysis Facility, QEII Medical Centre, The University of Western Australia, Crawley, WA, 6009, Australia |
| |
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. |
| |
Keywords: | Mechanosensitive channel of large conductance Green fluorescent protein Confocal microscopy Patch-clamp |
本文献已被 PubMed SpringerLink 等数据库收录! |
|