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Issue 15, 2012
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A novel microfluidics-based method for probing weak proteinprotein interactions

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Abstract

We report the use of a novel microfluidics-based method to detect weak proteinprotein interactions between membrane proteins. The tight junction protein, claudin-2, synthesised in vitro using a cell-free expression system in the presence of polymer vesicles as membrane scaffolds, was used as a model membrane protein. Individual claudin-2 molecules interact weakly, although the cumulative effect of these interactions is significant. This effect results in a transient decrease of average vesicle dispersivity and reduction in transport speed of claudin-2-functionalised vesicles. Polymer vesicles functionalised with claudin-2 were perfused through a microfluidic channel and the time taken to traverse a defined distance within the channel was measured. Functionalised vesicles took 1.19 to 1.69 times longer to traverse this distance than unfunctionalised ones. Coating the channel walls with protein A and incubating the vesicles with anti-claudin-2 antibodies prior to perfusion resulted in the functionalised vesicles taking 1.75 to 2.5 times longer to traverse this distance compared to the controls. The data show that our system is able to detect weak as well as strong proteinprotein interactions. This system offers researchers a portable, easily operated and customizable platform for the study of weak proteinprotein interactions, particularly between membrane proteins.

Graphical abstract: A novel microfluidics-based method for probing weak protein–protein interactions

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Publication details

The article was received on 05 Mar 2012, accepted on 03 Apr 2012 and first published on 04 Apr 2012


Article type: Paper
DOI: 10.1039/C2LC40228A
Citation: Lab Chip, 2012,12, 2726-2735
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    A novel microfluidics-based method for probing weak proteinprotein interactions

    D. C. Tan, I. P. M. Wijaya, M. Andreasson-Ochsner, E. N. Vasina, M. Nallani, W. Hunziker and E. Sinner, Lab Chip, 2012, 12, 2726
    DOI: 10.1039/C2LC40228A

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