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Studying mechanosensitive ion channels with an automated patch clamp
Authors:Maria Barthmes  Mac Donald F. Jose  Jan Peter Birkner  Andrea Brüggemann  Christian Wahl-Schott  Armağan Koçer
Affiliation:1. Nanion Technologies GmbH, Gabrielenstr. 9, 80636, Munich, Germany
4. Department Pharmazie, Pharmakologie für Naturwissenschaften, Ludwig-Maximilians-Universit?t München, Butenandtstr. 5-13, 81377, Munich, Germany
2. Department of Biochemistry, University of Groningen, Nijenborgh 4, 9747 AG, Groningen, The Netherlands
3. Single-Molecule Biophysics, University of Groningen, Nijenborgh 4, 9747 AG, Groningen, The Netherlands
Abstract:Patch clamp electrophysiology is the main technique to study mechanosensitive ion channels (MSCs), however, conventional patch clamping is laborious and success and output depends on the skills of the operator. Even though automated patch systems solve these problems for other ion channels, they could not be applied to MSCs. Here, we report on activation and single channel analysis of a bacterial mechanosensitive ion channel using an automated patch clamp system. With the automated system, we could patch not only giant unilamellar liposomes but also giant Escherichia coli (E. coli) spheroplasts. The tension sensitivity and channel kinetics data obtained in the automated system were in good agreement with that obtained from the conventional patch clamp. The findings will pave the way to high throughput fundamental and drug screening studies on mechanosensitive ion channels.
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