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A comparison of cysteine-conjugated nitroxide spin labels for pulse dipolar EPR spectroscopy
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dc.contributor.author | Ackermann, Katrin | |
dc.contributor.author | Chapman, Alexandra | |
dc.contributor.author | Bode, Bela E. | |
dc.date.accessioned | 2021-12-14T10:30:07Z | |
dc.date.available | 2021-12-14T10:30:07Z | |
dc.date.issued | 2021-12-13 | |
dc.identifier | 276954862 | |
dc.identifier | 036ecc73-d56b-47a2-b562-e1696e62465f | |
dc.identifier | 85121228597 | |
dc.identifier | 000736887400001 | |
dc.identifier.citation | Ackermann , K , Chapman , A & Bode , B E 2021 , ' A comparison of cysteine-conjugated nitroxide spin labels for pulse dipolar EPR spectroscopy ' , Molecules , vol. 26 , no. 24 , 7534 . https://doi.org/10.3390/molecules26247534 | en |
dc.identifier.issn | 1420-3049 | |
dc.identifier.other | ORCID: /0000-0002-3384-271X/work/105007038 | |
dc.identifier.uri | https://hdl.handle.net/10023/24508 | |
dc.description | This research was funded, in whole or in part, by the Wellcome Trust (099149/Z/12/Z and 204821/Z/16/Z). B.E.B. and K.A. acknowledge support from the Leverhulme Trust (RPG-2018–397). B.E.B. acknowledges equipment funding from BBSRC (BB/R013780/1 and BB/T017740/1). | en |
dc.description.abstract | The structure-function and materials paradigms drive research on the understanding of structures and structural heterogeneity of molecules and solids from materials science to structural biology. Functional insights into complex architectures are often gained from a suite of complementary physicochemical methods. In the context of biomacromolecular structures, the use of pulse dipolar electron paramagnetic resonance spectroscopy (PDS) has become increasingly popular. The main interest in PDS is providing long-range nanometre distance distributions that allow for identifying macromolecular topologies, validating structural models and conformational transitions as well as docking of quaternary complexes. Most commonly, cysteines are introduced into protein structures by site-directed mutagenesis and modified site-specifically to a spin-labelled side-chain such as a stable nitroxide radical. In this contribution, we investigate labelling by four different commercial labelling agents that react through different sulfur-specific reactions. Further, the distance distributions obtained are between spin-bearing moieties and need to be related to the protein structure via modelling approaches. Here, we compare two different approaches to modelling these distributions for all four side-chains. The results indicate that there are significant differences in the optimum labelling procedure. All four spin-labels show differences in the ease of labelling and purification. Further challenges arise from the different tether lengths and rotamers of spin-labelled side-chains; both influence the modelling and translation into structures. Our comparison indicates that the spin-label with the shortest tether in the spin-labelled side-group, (bis-(2,2,5,5-Tetramethyl-3-imidazoline-1-oxyl-4-yl) disulfide, may be underappreciated and could increase the resolution of structural studies by PDS if labelling conditions are optimised accordingly. | |
dc.format.extent | 16 | |
dc.format.extent | 23820797 | |
dc.language.iso | eng | |
dc.relation.ispartof | Molecules | en |
dc.subject | PDS | en |
dc.subject | PELDOR | en |
dc.subject | DEER | en |
dc.subject | Nitroxide spin label | en |
dc.subject | Comparative DEER analyzer | en |
dc.subject | mtsslSuite | en |
dc.subject | MMM | en |
dc.subject | QD Chemistry | en |
dc.subject | DAS | en |
dc.subject.lcc | QD | en |
dc.title | A comparison of cysteine-conjugated nitroxide spin labels for pulse dipolar EPR spectroscopy | en |
dc.type | Journal article | en |
dc.contributor.sponsor | BBSRC | en |
dc.contributor.sponsor | BBSRC | en |
dc.contributor.sponsor | The Leverhulme Trust | en |
dc.contributor.sponsor | The Wellcome Trust | en |
dc.contributor.sponsor | The Wellcome Trust | en |
dc.contributor.institution | University of St Andrews. School of Chemistry | en |
dc.contributor.institution | University of St Andrews. EaSTCHEM | en |
dc.contributor.institution | University of St Andrews. Biomedical Sciences Research Complex | en |
dc.contributor.institution | University of St Andrews. Centre of Magnetic Resonance | en |
dc.identifier.doi | 10.3390/molecules26247534 | |
dc.description.status | Peer reviewed | en |
dc.identifier.grantnumber | BB/R013780/1 | en |
dc.identifier.grantnumber | BB/T017740/1 | en |
dc.identifier.grantnumber | RPG-2018-397 | en |
dc.identifier.grantnumber | 099149/Z/12/Z | en |
dc.identifier.grantnumber | en |
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