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Issue 4, 2012
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Activity monitoring of functional OprM using a biomimetic microfluidic device

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Abstract

This paper describes the fabrication and use of a biomimetic microfluidic device for the monitoring of a functional porin reconstituted within a miniaturized suspended artificial bilayer lipid membrane (BLM). Such a microfluidic device allows for (1) fluidic and electrical access to both sides of the BLM and (2) reproducible membrane protein insertion and long-term electrical monitoring of its conductance (Gi), thanks to the miniaturization of the BLM. We demonstrate here for the first time the feasibility to insert a large trans-membrane protein through its β-barrel, and monitor its functional activity for more than 1 hour (limited by buffer evaporation). In this paper, we specifically used our device for the monitoring of OprM, a bacterial efflux channel involved in the multidrug resistance of the bacteria Pseudomonas aeruginosa. Sub-steps of the OprM channel conductance were detected during the electrical recordings within our device, which might be due to oscillations between several structural conformations (sub-states) adopted by the protein, as part of its opening mechanism. This work is a first step towards the establishment of a genuine platform dedicated to the investigation of bacterial proteins under reconstituted conditions, a very promising tool for the screening of new inhibitors against bacterial channels involved in drug resistance.

Graphical abstract: Activity monitoring of functional OprM using a biomimetic microfluidic device

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Article information


Submitted
24 Oct 2011
Accepted
02 Dec 2011
First published
04 Jan 2012

Analyst, 2012,137, 847-852
Article type
Paper

Activity monitoring of functional OprM using a biomimetic microfluidic device

W. Wang, L. Monlezun, M. Picard, P. Benas, O. Français, I. Broutin and B. Le Pioufle, Analyst, 2012, 137, 847
DOI: 10.1039/C2AN16007B

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