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Open Access High-spin Metal Centres in Dipolar EPR Spectroscopy

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The substitution of Mg2+ by Mn2+ in the bacterial DnaB helicase from Helicobacter pylori, an ATP:Mg2+-fuelled protein engine, allows electron paramagnetic resonance (EPR) spectroscopy to be performed on this system. EPR experiments make it possible to monitor nucleotide binding and to estimate the fraction of bound Mn2+ through relaxation measurements. Furthermore, by measuring spin–spin distances we probe the geometry within such multimeric assemblies using ultra-wideband double electron-electron resonance (DEER) and relaxation induced dipolar modulation enhancement (RIDME). The extraction of distance distributions from RIDME experiments on high-spin paramagnetic centres is influenced by the presence of dipolar frequency overtones. We show herein that we can correct for these overtones by using a modified kernel function in Tikhonov regularization analysis routines, and that the overtone coefficients for Mn2+ in the DnaB helicase are practically the same as in the previously studied Mn2+–Mn2+ model compounds.

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Keywords: DISTANCE MEASUREMENTS; EPR SPECTROSCOPY; GADOLINIUM; MANGANESE; MOTOR PROTEINS

Document Type: Research Article

Affiliations: 1: ETH Zurich, Laboratorium für Physikalische Chemie, Vladimir-Prelog-Weg 2, CH-8093 Zurich, Switzerland 2: Molecular Microbiology and Structural Biochemistry, Labex Ecofect UMR 5086 CNRS/Université de Lyon, Lyon, France 3: ETH Zurich, Laboratorium für Physikalische Chemie, Vladimir-Prelog-Weg 2, CH-8093 Zurich, Switzerland. [email protected]

Publication date: April 1, 2018

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  • International Journal for Chemistry and Official Membership Journal of the Swiss Chemical Society (SCS) and its Divisions

    CHIMIA, a scientific journal for chemistry in the broadest sense, is published 10 times a year and covers the interests of a wide and diverse readership. Contributions from all fields of chemistry and related areas are considered for publication in the form of Review Articles and Notes. A characteristic feature of CHIMIA are the thematic issues, each devoted to an area of great current significance.

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