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Magnetic Resonance An interactive open-access publication of the Groupement AMPERE
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https://doi.org/10.5194/mr-2020-15
© Author(s) 2020. This work is distributed under
the Creative Commons Attribution 4.0 License.
https://doi.org/10.5194/mr-2020-15
© Author(s) 2020. This work is distributed under
the Creative Commons Attribution 4.0 License.

  23 Jun 2020

23 Jun 2020

Review status
A revised version of this preprint is currently under review for the journal MR.

Orthogonally spin-labeled rulers help to identify crosstalk signals and improve DEER signal fidelity

Markus Teucher1, Mian Qi2, Ninive Cati2, Henrik Hintz2, Adelheid Godt2, and Enrica Bordignon1 Markus Teucher et al.
  • 1Faculty of Chemistry and Biochemistry, Ruhr University Bochum, Universitätsstraße 150, 44801 Bochum, Germany
  • 2Faculty of Chemistry and Center for Molecular Materials (CM2), Bielefeld University, Universitätsstraße 25, 33615 Bielefeld, Germany

Abstract. DEER spectroscopy applied to orthogonally spin-labeled proteins is a versatile technique which allows simplifying the assignment of distances in complex spin systems and thereby increasing the information content that can be obtained per sample. In fact, orthogonal spin labels can be independently addressed in DEER experiments due to spectroscopically non-overlapping central transitions, distinct relaxation times and/or transition moments. Here we focus on molecular rulers orthogonally labeled with nitroxide (NO) and gadolinium (Gd) spins, which give access to three distinct DEER channels, probing NO-NO, NO-Gd and Gd-Gd distances. It has been previously suggested that crosstalk signals between individual DEER channels might occur, for example, between NO and Gd due to their inevitable spectral overlap. However, a systematic study to address these issues has not yet been carried out. Here, we perform a thorough three-channel DEER analysis on mixtures of NO-NO, NO-Gd and Gd-Gd molecular rulers characterized by distinct, non-overlapping distance distributions to study under which conditions crosstalk signals occur and how they can be identified or suppressed to improve signal fidelity. This study will help to improve the assignment of the correct distances in homo- and hetero-complexes of orthogonally spin-labeled proteins.

Markus Teucher et al.

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Markus Teucher et al.

Markus Teucher et al.

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Latest update: 19 Sep 2020
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Short summary
With a pulsed dipolar EPR technique named Double Electron-Electron Resonance, we measure nanometer distances between spin labels attached to biomolecules. If more than one spin type is present (A and B), we can separately address AA, AB and BB distances via distinct spectroscopic channels, increasing the information content per sample. Here we investigate the appearance of unwanted channel crosstalks and suggest ways to identify and suppress them, thereby increasing signal fidelity.
With a pulsed dipolar EPR technique named Double Electron-Electron Resonance, we measure...
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