Orthogonal spin labeling using click chemistry for in vitro and in vivo applications

DC FieldValueLanguage
dc.contributor.authorKucher, Svetlana
dc.contributor.authorKorneev, Sergei
dc.contributor.authorTyagi, Swati
dc.contributor.authorApfelbaum, Ronja
dc.contributor.authorGrohmann, Dina
dc.contributor.authorLemke, Edward A.
dc.contributor.authorKlare, Johann P.
dc.contributor.authorSteinhoff, Heinz-Juergen
dc.contributor.authorKlose, Daniel
dc.date.accessioned2021-12-23T16:21:45Z-
dc.date.available2021-12-23T16:21:45Z-
dc.date.issued2017
dc.identifier.issn10907807
dc.identifier.urihttps://osnascholar.ub.uni-osnabrueck.de/handle/unios/14004-
dc.description.abstractSite-directed spin labeling for EPR- and NMR spectroscopy has mainly been achieved exploiting the specific reactivity of cysteines. For proteins with native cysteines or for in vivo applications, an alternative coupling strategy is required. In these cases click chemistry offers major benefits by providing a fast and highly selective, biocompatible reaction between azide and alkyne groups. Here, we establish click chemistry as a tool to target unnatural amino acids in vitro and in vivo using azide- and alkyne-functionalized spin labels. The approach is compatible with a variety of labels including reduction-sensitive nitroxides. Comparing spin labeling efficiencies from the copper-free with the strongly reducing copper(I)-catalyzed azide-alkyne click reaction, we find that the faster kinetics for the catalyzed reaction outrun reduction of the labile nitroxide spin labels and allow quantitative labeling yields within short reaction times. Inter spin distance measurements demonstrate that the novel side chain is suitable for paramagnetic NMR- or EPR-based conformational studies of macromolecular complexes. (C) 2016 Elsevier Inc. All rights reserved.
dc.description.sponsorshipDFGGerman Research Foundation (DFG)European Commission [SPP1601, SPP1623]; Emmy Noether program of the DFGGerman Research Foundation (DFG); Fond der Chemischen IndustrieFonds der Chemischen Industrie; Boehringer Ingelheim FondsBoehringer Ingelheim; We thank Christine Koehler for help with protein purification, Maxim Yulikov for assistance with the Q-Band DEER measurements and Chris Kay for reading the manuscript. H.J.S. & D.K. acknowledge funding from the DFG priority program SPP1601. E. A.L. thanks the Emmy Noether program and SPP1623 of the DFG for funding. D.G. acknowledges funding by the Fond der Chemischen Industrie and the Boehringer Ingelheim Fonds.
dc.language.isoen
dc.publisherACADEMIC PRESS INC ELSEVIER SCIENCE
dc.relation.ispartofJOURNAL OF MAGNETIC RESONANCE
dc.subject1,3-DIPOLAR CYCLOADDITIONS
dc.subjectAMINO-ACID
dc.subjectAZIDE-ALKYNE CYCLOADDITION
dc.subjectBiochemical Research Methods
dc.subjectBiochemistry & Molecular Biology
dc.subjectClick chemistry
dc.subjectDEER
dc.subjectDISTANCE MEASUREMENTS
dc.subjectEPR SPECTROSCOPY
dc.subjectESCHERICHIA-COLI
dc.subjectGENETIC-CODE
dc.subjectIn vivo EPR
dc.subjectLIVING CELLS
dc.subjectOrthogonal labeling
dc.subjectPhysics
dc.subjectPhysics, Atomic, Molecular & Chemical
dc.subjectPROTEINS
dc.subjectSite-directed spin labeling
dc.subjectSpectroscopy
dc.subjectTERMINAL ALKYNES
dc.titleOrthogonal spin labeling using click chemistry for in vitro and in vivo applications
dc.typejournal article
dc.identifier.doi10.1016/j.jmr.2016.12.001
dc.identifier.isiISI:000393533900005
dc.description.volume275
dc.description.startpage38
dc.description.endpage45
dc.contributor.orcid0000-0002-5888-0157
dc.contributor.orcid0000-0002-5761-5968
dc.contributor.orcid0000-0002-0634-0503
dc.contributor.orcid0000-0002-3244-906X
dc.contributor.orcid0000-0002-3597-0889
dc.contributor.orcid0000-0002-7522-9163
dc.contributor.researcheridH-3791-2014
dc.contributor.researcheridC-1428-2009
dc.contributor.researcheridJ-3337-2015
dc.identifier.eissn10960856
dc.publisher.place525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA
dcterms.isPartOf.abbreviationJ. Magn. Reson.
crisitem.author.deptFB 04 - Physik-
crisitem.author.deptFB 05 - Biologie/Chemie-
crisitem.author.deptUniversität Osnabrück-
crisitem.author.deptUniversität Osnabrück-
crisitem.author.deptUniversität Osnabrück-
crisitem.author.deptFB 04 - Physik-
crisitem.author.deptFB 04 - Physik-
crisitem.author.deptidfb04-
crisitem.author.deptidfb05-
crisitem.author.deptidfb04-
crisitem.author.deptidfb04-
crisitem.author.orcid0000-0002-3244-906X-
crisitem.author.orcid0000-0002-3597-0889-
crisitem.author.parentorgUniversität Osnabrück-
crisitem.author.parentorgUniversität Osnabrück-
crisitem.author.parentorgUniversität Osnabrück-
crisitem.author.parentorgUniversität Osnabrück-
crisitem.author.netidKuSv960-
crisitem.author.netidKoSe681-
crisitem.author.netidTySw001-
crisitem.author.netidApRo001-
crisitem.author.netidGrDi001-
crisitem.author.netidLeEd010-
crisitem.author.netidStHe633-
crisitem.author.netidKlDa004-
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