Two-Dimensional Trap for Ultrasensitive Quantification of Transient Protein Interactions
DC Element | Wert | Sprache |
---|---|---|
dc.contributor.author | Beutel, Oliver | |
dc.contributor.author | Roder, Friedrich | |
dc.contributor.author | Birkholz, Oliver | |
dc.contributor.author | Rickert, Christian | |
dc.contributor.author | Steinhoff, Heinz-Juergen | |
dc.contributor.author | Grzybek, Michal | |
dc.contributor.author | Coskun, Uenal | |
dc.contributor.author | Piehler, Jacob | |
dc.date.accessioned | 2021-12-23T16:19:13Z | - |
dc.date.available | 2021-12-23T16:19:13Z | - |
dc.date.issued | 2015 | |
dc.identifier.issn | 19360851 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/13041 | - |
dc.description.abstract | 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. | |
dc.description.sponsorship | Deutsche ForschungsgemeinschaftGerman Research Foundation (DFG) [SFB 944, TRR83]; German Federal Ministry of Education and Research grant; We thank G. Hikade and H. Kenneweg for technical support and A. Honigmann for fruitful discussions. The H6-tagged anti-GFP nanobody was kindly provided by U. Rothbauer, Tubingen, Germany. This project was supported by the Deutsche Forschungsgemeinschaft (Grant SFB 944 to J.P. and H.-J.S.; Grant TRR83 to U.C.) and by the German Federal Ministry of Education and Research grant to the German Center for Diabetes Research. | |
dc.language.iso | en | |
dc.publisher | AMER CHEMICAL SOC | |
dc.relation.ispartof | ACS NANO | |
dc.subject | ACTIVATION | |
dc.subject | BIMOLECULAR FLUORESCENCE COMPLEMENTATION | |
dc.subject | BINDING-SITE | |
dc.subject | Chemistry | |
dc.subject | Chemistry, Multidisciplinary | |
dc.subject | Chemistry, Physical | |
dc.subject | DOMAIN | |
dc.subject | EGF RECEPTOR | |
dc.subject | fluorescence microscopy | |
dc.subject | GROWTH-FACTOR RECEPTOR | |
dc.subject | lipid phase separation | |
dc.subject | LIPID-BILAYERS | |
dc.subject | LIVING CELLS | |
dc.subject | Materials Science | |
dc.subject | Materials Science, Multidisciplinary | |
dc.subject | Nanoscience & Nanotechnology | |
dc.subject | PLASMA-MEMBRANE | |
dc.subject | polymer-supported membrane | |
dc.subject | POLYMER-SUPPORTED MEMBRANES | |
dc.subject | protein-lipid interaction | |
dc.subject | protein-protein interaction | |
dc.subject | Science & Technology - Other Topics | |
dc.subject | signaling complexes | |
dc.title | Two-Dimensional Trap for Ultrasensitive Quantification of Transient Protein Interactions | |
dc.type | journal article | |
dc.identifier.doi | 10.1021/acsnano.5b02696 | |
dc.identifier.isi | ISI:000363915300033 | |
dc.description.volume | 9 | |
dc.description.issue | 10 | |
dc.description.startpage | 9783 | |
dc.description.endpage | 9791 | |
dc.contributor.orcid | 0000-0002-5888-0157 | |
dc.contributor.orcid | 0000-0003-4375-3144 | |
dc.contributor.orcid | 0000-0003-2083-0506 | |
dc.contributor.orcid | 0000-0001-6551-3219 | |
dc.contributor.researcherid | H-3791-2014 | |
dc.contributor.researcherid | F-4683-2011 | |
dc.contributor.researcherid | D-3802-2017 | |
dc.identifier.eissn | 1936086X | |
dc.publisher.place | 1155 16TH ST, NW, WASHINGTON, DC 20036 USA | |
dcterms.isPartOf.abbreviation | ACS Nano | |
crisitem.author.dept | FB 04 - Physik | - |
crisitem.author.dept | FB 05 - Biologie/Chemie | - |
crisitem.author.deptid | fb04 | - |
crisitem.author.deptid | fb05 | - |
crisitem.author.orcid | 0000-0002-2143-2270 | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.netid | StHe633 | - |
crisitem.author.netid | PiJa938 | - |
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geprüft am 23.05.2024