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MscL - "Lipid bilayer recordings of a mechanosensitive channel, MscL, using Nanion’s pressure clampchannel, MscL, using Nanion’s pressure clamp"

icon pap   Port-a-Patch and    icon vpp   Vesicle Prep Pro application note   logo pdf   (0.3 MB)


Solvent-free planar lipid bilayers were formed in an automated manner using suction to attract a giant unilamellar vesicle (GUV) to the patch clamp chip which subsequently bursts across the aperture. Incubation of GUVs with purified MscL channel protein yielded proteoliposomes. These proteoliposomes allow for immediate recording of channel activity after GUV sealing. The rapid formation of protein-containing planar lipid bilayers is of potential use for the efficient electrophysiological characterization of MscL as shown here and also other ion channel proteins of interest.
In order to study the effect of pressure, the functional MscL purified was reconstituted in our system. The reconstitution was done in GUVs and then bilayers were formed on a chip (Kreir, Farre et al. 2008). The Port-a-Patch system has a pump controlled by a computer and could apply from +300 to -300 mBar and is controlled via software allowing accurate pressure control. All pressure applications could be visualized and recorded at the same time as the recordings.


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