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Beutel, O.* ; Roder, F.* ; Birkholz, O.* ; Rickert, C.* ; Steinhoff, H.* ; Grzybek, M. ; Coskun, Ü. ; Piehler, J.*

Two-dimensional trap for ultrasensitive quantification of transient protein interactions.

ACS Nano 9, 9783-9791 (2015)
DOI
Open Access Green as soon as Postprint is submitted to ZB.
We present an ultrasensitive technique for quantitative protein–protein interaction analysis in a two-dimensional format based on phase-separated, micropatterned membranes. Interactions between proteins captured to lipid probes via an affinity tag trigger partitioning into the liquid-ordered phase, which is readily quantified by fluorescence imaging. Based on a calibration with well-defined low-affinity protein–protein interactions, equilibrium dissociation constants >1 mM were quantified. Direct capturing of proteins from mammalian cell lysates enabled us to detect homo- and heterodimerization of signal transducer and activator of transcription proteins. Using the epidermal growth factor receptor (EGFR) as a model system, quantification of low-affinity interactions between different receptor domains contributing to EGFR dimerization was achieved. By exploitation of specific features of the membrane-based assay, the regulation of EGFR dimerization by lipids was demonstrated.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Protein-protein Interaction ; Polymer-supported Membrane ; Lipid Phase Separation ; Fluorescence Microscopy ; Signaling Complexes ; Protein-lipid Interaction
Language english
Publication Year 2015
HGF-reported in Year 2015
ISSN (print) / ISBN 1936-0851
e-ISSN 1936-086X
Journal ACS Nano
Quellenangaben Volume: 9, Issue: 10, Pages: 9783-9791 Article Number: , Supplement: ,
Publisher American Chemical Society (ACS)
Reviewing status Peer reviewed
Institute(s) Institute of Pancreatic Islet Research (IPI)
POF-Topic(s) 90000 - German Center for Diabetes Research
Research field(s) Helmholtz Diabetes Center
PSP Element(s) G-502600-002
DOI 10.1021/acsnano.5b02696
WOS ID WOS:000363915300033
Erfassungsdatum 2015-11-27
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