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Small Gd(III) Tags for Gd(III)-Gd(III) Distance Measurements in Proteins by EPR Spectroscopy

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dc.contributor.authorProkopiou, Georgia
dc.contributor.authorLee, Michael D.
dc.contributor.authorCollauto, A.
dc.contributor.authorAbdelkader, Elwy
dc.contributor.authorBahrenberg, Thorsten
dc.contributor.authorFeintuch, Akiva
dc.contributor.authorRamirez Cohen, Marie
dc.contributor.authorClayton, Jessica
dc.contributor.authorSwarbrick, James
dc.contributor.authorGraham, Bim
dc.contributor.authorOtting, Gottfried
dc.contributor.authorGoldfarb, Daniella
dc.date.accessioned2019-08-12T04:41:03Z
dc.date.issued2018
dc.date.updated2022-08-07T08:16:15Z
dc.description.abstractThe C7-Gd and C8-Gd tags are compact hydrophilic cyclen-based lanthanide tags for conjugation to cysteine residues in proteins. The tags are enantiomers, which differ in the configuration of the 2-hydroxylpropyl pendant arms coordinating the lanthanide ion. Here, we report the electron paramagnetic resonance (EPR) performance of the C7-Gd (S configuration) and C8-Gd (R configuration) tags loaded with Gd(III) on two mutants of the homodimeric ERp29 protein. The W-band EPR spectra were found to differ between the tags in the free state and after conjugation to the protein. In addition, the spectra were sensitive to the labeling position, which may originate from an environment-dependent charge density on the Gd(III)-coordinating oxygens. This is in agreement with previous NMR experiments with different lanthanide ions, which suggested sensitivity to H-bonding. W-band 1H-ENDOR (electron–electron double resonance) experiments detected effects from orientation selection in the central transition, due to a relatively narrow distribution in the ZFS parameters as indicated by simulations. In contrast, the distance distributions derived from DEER (double electron–electron resonance) measurements were insensitive to the R or S configuration of the tags and did not exhibit any orientation selection effects. The DEER measurements faithfully reflected the different widths of the distance distributions at the different protein sites in agreement with previous DEER measurements using other Gd(III) tags. Due to their small size, short tether to the protein, and a broad central EPR transition, the C7-Gd and C8-Gd tags are attractive Gd(III) tags for measurements of relatively short (<4 nm) distances by EPR spectroscopy.
dc.description.sponsorshipD.G. acknowledges the support of the Israel Science Foundation (ISF grant No. 334/ 14). J.C. was supported by NSF grant MCB 1617025. This research was made possible in part by the historic generosity of the Harold Perlman Family (D.G.). D.G. holds the Erich Klieger Professorial Chair in Chemical Physics. T.B. acknowledges financial support from the Minerva Foundation. G.O. thanks the Australian Research Council (ARC) for a Laureate Fellowship, B.G. thanks the ARC for a Future Fellowship, and G.O. and B.G. thank the ARC for financial support.en_AU
dc.format.mimetypeapplication/pdfen_AU
dc.identifier.issn0020-1669en_AU
dc.identifier.urihttp://hdl.handle.net/1885/164995
dc.language.isoen_AUen_AU
dc.provenancehttps://v2.sherpa.ac.uk/id/publication/7780..."The Accepted Version can be archived in a non-commercial institutional repository if required by funder. 12 months embargo." from SHERPA/RoMEO site (as at 2/09/2020)." This document is the Accepted Manuscript version of a Published Work that appeared in final form in Inorganic Chemistry, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://dx.doi.org/10.1021/acs.inorgchem.8b00133"
dc.publisherAmerican Chemical Society
dc.rights© 2018 American Chemical Society
dc.sourceInorganic Chemistry
dc.titleSmall Gd(III) Tags for Gd(III)-Gd(III) Distance Measurements in Proteins by EPR Spectroscopy
dc.typeJournal article
dcterms.accessRightsOpen Access
local.bibliographicCitation.issue9en_AU
local.bibliographicCitation.lastpage5059en_AU
local.bibliographicCitation.startpage5048en_AU
local.contributor.affiliationProkopiou, Georgia, Department of Chemical Physicsen_AU
local.contributor.affiliationLee, Michael D., Monash Universityen_AU
local.contributor.affiliationCollauto, A., Weizmann Institute of Scienceen_AU
local.contributor.affiliationAbdelkader, Elwy, College of Science, ANUen_AU
local.contributor.affiliationBahrenberg, Thorsten, Department of Chemical Physicsen_AU
local.contributor.affiliationFeintuch, Akiva, Weizmann Institute of Scienceen_AU
local.contributor.affiliationRamirez Cohen, Marie, Department of Chemical Physicsen_AU
local.contributor.affiliationClayton, Jessica, Department of Physicsen_AU
local.contributor.affiliationSwarbrick, James, Monash Universityen_AU
local.contributor.affiliationGraham, Bim, Monash Institute of Pharmaceutical Sciencesen_AU
local.contributor.affiliationOtting, Gottfried, College of Science, ANUen_AU
local.contributor.affiliationGoldfarb, Daniella, Weizmann Institute of Scienceen_AU
local.contributor.authoremailrepository.admin@anu.edu.auen_AU
local.contributor.authoruidAbdelkader, Elwy, u5154721en_AU
local.contributor.authoruidOtting, Gottfried, u4046684en_AU
local.description.notesImported from ARIESen_AU
local.identifier.absfor060112 - Structural Biology (incl. Macromolecular Modelling)en_AU
local.identifier.absfor030503 - Organic Chemical Synthesisen_AU
local.identifier.absseo970106 - Expanding Knowledge in the Biological Sciencesen_AU
local.identifier.ariespublicationa383154xPUB10011en_AU
local.identifier.citationvolume57en_AU
local.identifier.doi10.1021/acs.inorgchem.8b00133en_AU
local.identifier.scopusID2-s2.0-85046653179
local.identifier.thomsonIDWOS:000431833500035
local.identifier.uidSubmittedBya383154en_AU
local.publisher.urlhttps://pubs.acs.org/en_AU
local.type.statusAccepted Versionen_AU

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