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dc.contributor.authorHu, Shu-Hong
dc.contributor.authorWhitten, Andrew E
dc.contributor.authorKing, Gordon J
dc.contributor.authorJones, Alun
dc.contributor.authorRowland, Alexander F
dc.contributor.authorJames, David E
dc.contributor.authorMartin, Jennifer L
dc.date.accessioned2018-01-19T04:58:31Z
dc.date.available2018-01-19T04:58:31Z
dc.date.issued2012
dc.identifier.issn1932-6203
dc.identifier.doi10.1371/journal.pone.0041731
dc.identifier.urihttp://hdl.handle.net/10072/171989
dc.description.abstractARHGAP22 is a RhoGAP protein comprising an N-terminal PH domain, a RhoGAP domain and a C-terminal coiled-coil domain. It has recently been identified as an Akt substrate that binds 14-3-3 proteins in response to treatment with growth factors involved in cell migration. We used a range of biophysical techniques to investigate the weak interaction between 14-3-3 and a truncated form of ARHGAP22 lacking the coiled-coil domain. This weak interaction could be stabilized by chemical cross-linking which we used to show that: a monomer of ARHGAP22 binds a dimer of 14-3-3; the ARHGAP22 PH domain is required for the 14-3-3 interaction; the RhoGAP domain is unlikely to participate in the interaction; Ser16 is the more important of two predicted 14-3-3 binding sites; and, phosphorylation of Ser16 may not be necessary for 14-3-3 interaction under the conditions we used. Small angle X-ray scattering and cross-link information were used to generate solution structures of the isolated proteins and of the cross-linked ARHGAP22:14-3-3 complex, showing that no major rearrangement occurs in either protein upon binding, and supporting a role for the PH domain and N-terminal peptide of ARHGAP22 in the 14-3-3 interaction. Small-angle X-ray scattering measurements of mixtures of ARHGAP22 and 14-3-3 were used to establish that the affinity of the interaction is ,30 mM.
dc.description.peerreviewedYes
dc.languageEnglish
dc.publisherPublic Library of Sciences
dc.relation.ispartofpagefrome41731-1
dc.relation.ispartofpagetoe41731-11
dc.relation.ispartofissue8
dc.relation.ispartofjournalPLoS One
dc.relation.ispartofvolume7
dc.subject.fieldofresearchBiochemistry and Cell Biology not elsewhere classified
dc.subject.fieldofresearchcode060199
dc.titleThe Weak Complex between RhoGAP Protein ARHGAP22 and Signal Regulatory Protein 14-3-3 Has 1:2 Stoichiometry and a Single Peptide Binding Mode
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
dcterms.licensehttps://creativecommons.org/licenses/by/4.0/
dc.description.versionPublished
gro.rights.copyright© 2012 Hu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
gro.hasfulltextFull Text
gro.griffith.authorMartin, Jennifer
gro.griffith.authorKing, Gordon


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