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Novel, highly-sensitive platform to detect membrane transport activity

Detection sensitivity enhanced up to 6 orders of magnitude

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Graduate School of Engineering / Faculty of Engineering
2014/08/04

The research group of Assistant Professor Rikiya Watanabe and Professor Hiroyuki Noji at the University of Tokyo’s Graduate School of Engineering have developed a novel platform to measure membrane transport activity with extremely high sensitivity enhanced up to 6 orders of magnitude over conventional approaches.

Measurement of membrane transport activity using novel lipid membrane chip. (Left) Image of novel lipid membrane chip. (Right) Measurement of transport activity using a fluorescent indicator. The fluorescence intensity difference depends on activity level.

© 2014 Rikiya Watanabe.
Measurement of membrane transport activity using novel lipid membrane chip. (Left) Image of novel lipid membrane chip. (Right) Measurement of transport activity using a fluorescent indicator. The fluorescence intensity difference depends on activity level.

Membrane proteins, which are located on the cell membrane, play various pivotal roles in cell functions including signal transduction and energy production. Most commercially-available medicines target membrane proteins, and in recent years research has focused on membrane transporters, a group of membrane proteins that transport substrate molecules across the cell membrane and are an optimal pharmacological target due to their physiological importance. Although quantitatively measuring the transport activity of transporters is essential to evaluate the drug efficacy, it remains difficult as current methods have low detection sensitivity and limited target applicability.

To address these issues, the research group developed novel technologies including 1) high throughput formation of artificial lipid bilayer membranes with high stability and high compatibility to membrane proteins; 2) a custom-made lipid membrane chip that has more than 100,000 micro-chambers sealed with artificial lipid bilayers; 3) a method for the highly sensitive detection of membrane transporter transport activity. Using these technologies, they enhanced the detection sensitivity of transporter activity up to 6 orders of magnitude over conventional approaches, allowing single-molecule analysis of various membrane transporters, e.g. FoF1-ATP synthase, and α-hemolysin.

It is expected that the lipid membrane chip developed in this research will pave the way for novel analytical and pharmacological applications, e.g. the development of efficient drug screening platforms that target membrane transporters.

Press release

Paper

Rikiya Watanabe, Naoki Soga, Daishi Fujita, Kazuhito V Tabata, Lisa Yamauchi, Soo Hyeon Kim, Daisuke Asanuma, Mako Kamiya, Yasuteru Urano, Hiroaki Suga, & Hiroyuki Noji,
“Arrayed Lipid Bilayer Chambers Allow Single-Molecule Analysis of Membrane Transporter Activity”,
Nature Communications 2014, 5, 4519, doi: 10.1038/ncomms5519.
Article link (Publication, UTokyo Repository)

Links

Graduate School of Engineering

Department of Applied Chemistry, Graduate School of Engineering

Noji Laboratory, Department of Applied Chemistry, Graduate School of Engineering (Japanese)

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