Tracking Conformational Changes in Calmodulin in vitro, in Cell Extract, and in Cells by Electron Paramagnetic Resonance Distance Measurements
dc.contributor.author | Dalaloyan, Arina | |
dc.contributor.author | Martorana, Andrea | |
dc.contributor.author | Barak, Yoav | |
dc.contributor.author | Gataulin, Diana | |
dc.contributor.author | Reuveny, Eitan | |
dc.contributor.author | Howe, Andrew | |
dc.contributor.author | Elbaum, Michael | |
dc.contributor.author | Albeck, Shira | |
dc.contributor.author | Unger, Tamar | |
dc.contributor.author | Frydman, Veronica | |
dc.contributor.author | Abdelkader, Elwy | |
dc.contributor.author | Otting, Gottfried | |
dc.date.accessioned | 2020-03-12T04:57:59Z | |
dc.date.issued | 2019 | |
dc.date.updated | 2019-11-25T07:40:50Z | |
dc.description.abstract | It 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.sponsorship | D. 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.mimetype | application/pdf | en_AU |
dc.identifier.issn | 1439-4235 | en_AU |
dc.identifier.uri | http://hdl.handle.net/1885/202156 | |
dc.language.iso | en_AU | en_AU |
dc.publisher | Wiley | en_AU |
dc.rights | © 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim | en_AU |
dc.source | ChemPhysChem | en_AU |
dc.title | Tracking Conformational Changes in Calmodulin in vitro, in Cell Extract, and in Cells by Electron Paramagnetic Resonance Distance Measurements | en_AU |
dc.type | Journal article | en_AU |
local.bibliographicCitation.issue | 14 | en_AU |
local.bibliographicCitation.lastpage | 1868 | en_AU |
local.bibliographicCitation.startpage | 1860 | en_AU |
local.contributor.affiliation | Dalaloyan, Arina, The Weizmann Institute of Science | en_AU |
local.contributor.affiliation | Martorana, Andrea, The Weizmann Institute of Science | en_AU |
local.contributor.affiliation | Barak, Yoav, The Weizmann Institute of Science | en_AU |
local.contributor.affiliation | Gataulin, Diana, The Weizmann Institute of Science | en_AU |
local.contributor.affiliation | Reuveny, Eitan, The Weizmann Institute of Science | en_AU |
local.contributor.affiliation | Howe, Andrew, The Weizmann Institute of Science | en_AU |
local.contributor.affiliation | Elbaum, Michael, The Weizmann Institute of Science | en_AU |
local.contributor.affiliation | Albeck, Shira, Weizmann Institute of Science | en_AU |
local.contributor.affiliation | Unger, Tamar, The Weizmann Institute of Science | en_AU |
local.contributor.affiliation | Frydman, Veronica, Weizmann Institute of Science | en_AU |
local.contributor.affiliation | Abdelkader, Elwy, College of Science, ANU | en_AU |
local.contributor.affiliation | Otting, Gottfried, College of Science, ANU | en_AU |
local.contributor.authoremail | repository.admin@anu.edu.au | en_AU |
local.contributor.authoruid | Abdelkader, Elwy, u5154721 | en_AU |
local.contributor.authoruid | Otting, Gottfried, u4046684 | en_AU |
local.description.embargo | 2037-12-31 | |
local.description.notes | Imported from ARIES | en_AU |
local.identifier.absfor | 030403 - Characterisation of Biological Macromolecules | en_AU |
local.identifier.absfor | 030406 - Proteins and Peptides | en_AU |
local.identifier.absfor | 030606 - Structural Chemistry and Spectroscopy | en_AU |
local.identifier.absseo | 970103 - Expanding Knowledge in the Chemical Sciences | en_AU |
local.identifier.ariespublication | u3102795xPUB4172 | en_AU |
local.identifier.citationvolume | 20 | en_AU |
local.identifier.doi | 10.1002/cphc.201900341 | en_AU |
local.identifier.scopusID | 2-s2.0-85068157783 | |
local.identifier.uidSubmittedBy | u3102795 | en_AU |
local.publisher.url | https://www.wiley.com/en-gb | en_AU |
local.type.status | Published Version | en_AU |
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