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dc.contributor.authorOh, Dongmyung
dc.contributor.authorOgiue-Ikeda, Mari
dc.contributor.authorJia, Lin
dc.contributor.authorMachida, Kazuya
dc.contributor.authorYu, Ji
dc.contributor.authorJadwin, Joshua A.
dc.contributor.authorCurran, Timothy G.
dc.contributor.authorWhite, Forest M.
dc.contributor.authorMayer, Bruce J.
dc.date.accessioned2016-05-23T15:38:50Z
dc.date.available2016-05-23T15:38:50Z
dc.date.issued2016-04
dc.date.submitted2015-09
dc.identifier.issn2050-084X
dc.identifier.urihttp://hdl.handle.net/1721.1/102619
dc.description.abstractWhile the affinities and specificities of SH2 domain-phosphotyrosine interactions have been well characterized, spatio-temporal changes in phosphosite availability in response to signals, and their impact on recruitment of SH2-containing proteins in vivo, are not well understood. To address this issue, we used three complementary experimental approaches to monitor phosphorylation and SH2 binding in human A431 cells stimulated with epidermal growth factor (EGF): 1) phospho-specific mass spectrometry; 2) far-Western blotting; and 3) live cell single-molecule imaging of SH2 membrane recruitment. Far-Western and MS analyses identified both well-established and previously undocumented EGF-dependent tyrosine phosphorylation and binding events, as well as dynamic changes in binding patterns over time. In comparing SH2 binding site phosphorylation with SH2 domain membrane recruitment in living cells, we found in vivo binding to be much slower. Delayed SH2 domain recruitment correlated with clustering of SH2 domain binding sites on the membrane, consistent with membrane retention via SH2 rebinding.en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (Grant U01CA154966)en_US
dc.language.isoen_US
dc.publishereLife Sciences Publications, Ltd.en_US
dc.relation.isversionofhttp://dx.doi.org/10.7554/eLife.11835en_US
dc.rightsCreative Commons Attributionen_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en_US
dc.sourceeLife Sciences Publications, Ltd.en_US
dc.titleTime-resolved multimodal analysis of Src Homology 2 (SH2) domain binding in signaling by receptor tyrosine kinasesen_US
dc.typeArticleen_US
dc.identifier.citationJadwin, Joshua A, Dongmyung Oh, Timothy G Curran, Mari Ogiue-Ikeda, Lin Jia, Forest M White, Kazuya Machida, Ji Yu, and Bruce J Mayer. “Time-Resolved Multimodal Analysis of Src Homology 2 (SH2) Domain Binding in Signaling by Receptor Tyrosine Kinases.” eLife 5 (April 13, 2016).en_US
dc.contributor.departmentDavid H. Koch Institute for Integrative Cancer Research at MITen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biological Engineeringen_US
dc.contributor.mitauthorCurran, Timothy G.en_US
dc.contributor.mitauthorWhite, Forest M.en_US
dc.relation.journaleLifeen_US
dc.eprint.versionFinal published versionen_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dspace.orderedauthorsJadwin, Joshua A; Oh, Dongmyung; Curran, Timothy G; Ogiue-Ikeda, Mari; Jia, Lin; White, Forest M; Machida, Kazuya; Yu, Ji; Mayer, Bruce Jen_US
dspace.embargo.termsNen_US
dc.identifier.orcidhttps://orcid.org/0000-0002-1545-1651
mit.licenseOPEN_ACCESS_POLICYen_US


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