Real-time single-molecule co-immunoprecipitation analyses reveal cancer-specific Ras signalling dynamics

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dc.contributor.authorH W Lee-
dc.contributor.authorT Kyung-
dc.contributor.authorJ Yoo-
dc.contributor.authorT Kim-
dc.contributor.authorC Chung-
dc.contributor.authorJ Y Ryu-
dc.contributor.authorHanki Lee-
dc.contributor.authorK Park-
dc.contributor.authorS Lee-
dc.contributor.authorW D Jones-
dc.contributor.authorD S Lim-
dc.contributor.authorC Hyeon-
dc.contributor.authorW D Heo-
dc.contributor.authorT Y Yoon-
dc.date.accessioned2017-04-19T09:46:15Z-
dc.date.available2017-04-19T09:46:15Z-
dc.date.issued2013-
dc.identifier.issn20411723-
dc.identifier.uri10.1038/ncomms2507ko
dc.identifier.urihttps://oak.kribb.re.kr/handle/201005/11655-
dc.description.abstractCo-immunoprecipitation (co-IP) has become a standard technique, but its protein-band output provides only static, qualitative information about protein-protein interactions. Here we demonstrate a real-time single-molecule co-IP technique that generates real-time videos of individual protein-protein interactions as they occur in unpurified cell extracts. By analysing single Ras-Raf interactions with a 50-ms time resolution, we have observed transient intermediates of the protein-protein interaction and determined all the essential kinetic rates. Using this technique, we have quantified the active fraction of native Ras proteins in xenograft tumours, normal tissue and cancer cell lines. We demonstrate that the oncogenic Ras mutations selectively increase the active-Ras fraction by one order of magnitude, without affecting total Ras levels or single-molecule signalling kinetics. Our approach allows us to probe the previously hidden, dynamic aspects of weak protein-protein interactions. It also suggests a path forward towards precision molecular diagnostics at the protein-protein interaction level.-
dc.publisherSpringer-Nature Pub Group-
dc.titleReal-time single-molecule co-immunoprecipitation analyses reveal cancer-specific Ras signalling dynamics-
dc.title.alternativeReal-time single-molecule co-immunoprecipitation analyses reveal cancer-specific Ras signalling dynamics-
dc.typeArticle-
dc.citation.titleNature Communications-
dc.citation.number0-
dc.citation.endPage1505-
dc.citation.startPage1505-
dc.citation.volume4-
dc.contributor.alternativeName이홍원-
dc.contributor.alternativeName경태윤-
dc.contributor.alternativeName유장현-
dc.contributor.alternativeName김택훈-
dc.contributor.alternativeName정차욱-
dc.contributor.alternativeName류지영-
dc.contributor.alternativeName이한기-
dc.contributor.alternativeName박기현-
dc.contributor.alternativeName이상규-
dc.contributor.alternativeNameJones-
dc.contributor.alternativeName임대식-
dc.contributor.alternativeName현창봉-
dc.contributor.alternativeName허원도-
dc.contributor.alternativeName윤태영-
dc.identifier.bibliographicCitationNature Communications, vol. 4, pp. 1505-1505-
dc.identifier.doi10.1038/ncomms2507-
dc.description.journalClassY-
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