Tracking Conformational Changes in Calmodulin in vitro, in Cell Extract, and in Cells by Electron Paramagnetic Resonance Distance Measurements

dc.contributor.authorDalaloyan, Arina
dc.contributor.authorMartorana, Andrea
dc.contributor.authorBarak, Yoav
dc.contributor.authorGataulin, Diana
dc.contributor.authorReuveny, Eitan
dc.contributor.authorHowe, Andrew
dc.contributor.authorElbaum, Michael
dc.contributor.authorAlbeck, Shira
dc.contributor.authorUnger, Tamar
dc.contributor.authorFrydman, Veronica
dc.contributor.authorAbdelkader, Elwy
dc.contributor.authorOtting, Gottfried
dc.date.accessioned2020-03-12T04:57:59Z
dc.date.issued2019
dc.date.updated2019-11-25T07:40:50Z
dc.description.abstractIt is an open question whether the conformations of proteins sampled in dilute solutions are the same as in the cellular environment. Here we address this question by double electron‐electron resonance (DEER) distance measurements with Gd(III) spin labels to probe the conformations of calmodulin (CaM) in vitro, in cell extract, and in human HeLa cells. Using the CaM mutants N53C/T110C and T34C/T117C labeled with maleimide‐DOTA‐Gd(III) in the N‐ and C‐terminal domains, we observed broad and varied interdomain distance distributions. The in vitro distance distributions of apo‐CaM and holo‐CaM in the presence and absence of the IQ target peptide can be described by combinations of closed, open, and collapsed conformations. In cell extract, apo‐ and holo‐CaM bind to target proteins in a similar way as apo‐ and holo‐CaM bind to IQ peptide in vitro. In HeLa cells, however, in the presence or absence of elevated in‐cell Ca2+ levels CaM unexpectedly produced more open conformations and very broad distance distributions indicative of many different interactions with in‐cell components. These results show‐case the importance of in‐cell analyses of protein structures.en_AU
dc.description.sponsorshipD. G. acknowledges the support of the Israel Science Foundation (ISF grant No. 334/14). D. G. holds the Erich Klieger Professorial Chair in Chemical Physics. G. O. thanks the Australian Research Council for financial support, including a Laureate Fellowship. In addition, this work was supported in part by the Israel Science Foundation ( grant NO. 1248/15) to E.R.en_AU
dc.format.mimetypeapplication/pdfen_AU
dc.identifier.issn1439-4235en_AU
dc.identifier.urihttp://hdl.handle.net/1885/202156
dc.language.isoen_AUen_AU
dc.publisherWileyen_AU
dc.rights© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheimen_AU
dc.sourceChemPhysChemen_AU
dc.titleTracking Conformational Changes in Calmodulin in vitro, in Cell Extract, and in Cells by Electron Paramagnetic Resonance Distance Measurementsen_AU
dc.typeJournal articleen_AU
local.bibliographicCitation.issue14en_AU
local.bibliographicCitation.lastpage1868en_AU
local.bibliographicCitation.startpage1860en_AU
local.contributor.affiliationDalaloyan, Arina, The Weizmann Institute of Scienceen_AU
local.contributor.affiliationMartorana, Andrea, The Weizmann Institute of Scienceen_AU
local.contributor.affiliationBarak, Yoav, The Weizmann Institute of Scienceen_AU
local.contributor.affiliationGataulin, Diana, The Weizmann Institute of Scienceen_AU
local.contributor.affiliationReuveny, Eitan, The Weizmann Institute of Scienceen_AU
local.contributor.affiliationHowe, Andrew, The Weizmann Institute of Scienceen_AU
local.contributor.affiliationElbaum, Michael, The Weizmann Institute of Scienceen_AU
local.contributor.affiliationAlbeck, Shira, Weizmann Institute of Scienceen_AU
local.contributor.affiliationUnger, Tamar, The Weizmann Institute of Scienceen_AU
local.contributor.affiliationFrydman, Veronica, Weizmann Institute of Scienceen_AU
local.contributor.affiliationAbdelkader, Elwy, College of Science, ANUen_AU
local.contributor.affiliationOtting, Gottfried, College of Science, ANUen_AU
local.contributor.authoremailrepository.admin@anu.edu.auen_AU
local.contributor.authoruidAbdelkader, Elwy, u5154721en_AU
local.contributor.authoruidOtting, Gottfried, u4046684en_AU
local.description.embargo2037-12-31
local.description.notesImported from ARIESen_AU
local.identifier.absfor030403 - Characterisation of Biological Macromoleculesen_AU
local.identifier.absfor030406 - Proteins and Peptidesen_AU
local.identifier.absfor030606 - Structural Chemistry and Spectroscopyen_AU
local.identifier.absseo970103 - Expanding Knowledge in the Chemical Sciencesen_AU
local.identifier.ariespublicationu3102795xPUB4172en_AU
local.identifier.citationvolume20en_AU
local.identifier.doi10.1002/cphc.201900341en_AU
local.identifier.scopusID2-s2.0-85068157783
local.identifier.uidSubmittedByu3102795en_AU
local.publisher.urlhttps://www.wiley.com/en-gben_AU
local.type.statusPublished Versionen_AU

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