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Quantitative time-resolved measurement of membrane protein–ligand interactions using microcantilever array sensors

Braun, Thomas and Ghatkesar, Murali Krishna and Backmann, Natalija and Grange, Wilfried and Boulanger, Pascale and Letellier, Lucienne and Lang, Hans-Peter and Bietsch, Alex and Gerber, Christoph and Hegner, Martin (2009) Quantitative time-resolved measurement of membrane protein–ligand interactions using microcantilever array sensors. Nature Nanotechnology, 4 (3). pp. 179-185. ISSN 1748-3387.

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Membrane proteins are central to many biological processes, and the interactions between transmembrane protein receptors and their ligands are of fundamental importance in medical research. However, measuring and characterizing these interactions is challenging. Here we report that sensors based on arrays of resonating microcantilevers can measure such interactions under physiological conditions. A protein receptor — the FhuA receptor of Escherichia coli — is crystallized in liposomes, and the proteoliposomes then immobilized on the chemically activated gold-coated surface of the sensor by ink-jet spotting in a humid environment, thus keeping the receptors functional. Quantitative mass-binding measurements of the bacterial virus T5 at subpicomolar concentrations are performed. These experiments demonstrate the potential of resonating microcantilevers for the specific, label-free and time-resolved detection of membrane protein–ligand interactions in a micro-array format.

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Additional Information:© 2009 Macmillan Publishers Limited. Published online: 18 January 2009. We thank A. Engel for the use of facilities for protein reconstitution at the Biozentrum, University of Basel, and M. Chami for fruitful discussions. We thank D. Mathys, M. Du¨ggelin and M. Du¨rrenberger for their help with the SEM analysis. Financial and general support is acknowledged from Swiss National Science Foundation (NCCR Nanoscale Science), the Commission for Technology and Innovation (CTI) (TOPNANO21), the European Learning and Teaching Mobility Regio Network, the G.H. Endress Foundation, the Novartis Foundation and the Science Foundation Ireland CSET programme. M.K.G. thanks the Swiss National Science Foundation and the Novartis Foundation for a research fellowship. T.B. and M.H. conceived and designed the experiments; T.B. and M.K.G. performed the experiments; T.B., M.K.G., W.G. and M.H. analysed the data; P.B. and L.L. contributed expertise in membrane proteins and phages; A.B. and T.B. were responsible for the ink-jet spotting of the membrane proteins; T.B., N.B., W.G., H.P.L. and M.H. co-wrote the paper. All authors discussed the results and commented on the manuscript.
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Swiss National Science FoundationUNSPECIFIED
Commission for Technology and Innovation (CTI)TOPNANO21
European Learning and Teaching Mobility Regio NetworkUNSPECIFIED
G.H. Endress FoundationUNSPECIFIED
Novartis FoundationUNSPECIFIED
Science Foundation IrelandUNSPECIFIED
Issue or Number:3
Record Number:CaltechAUTHORS:20090514-145726853
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:14220
Deposited By: Ruth Sustaita
Deposited On:14 Aug 2009 15:12
Last Modified:03 Oct 2019 00:48

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