2'-Alkynyl spin-labelling is a minimally perturbing tool for DNA structural analysis

Hardwick JS, Haugland MM, El-Sagheer AH, Ptchelkine D, Beierlein F, Lane AN, Brown T, Lovett JE, Anderson EA (2020)

Publication Type: Journal article

Publication year: 2020

Journal

Nucleic Acids Research Oxford University Press

Book Volume: 48

Pages Range: 2830-2840

Journal Issue: 6

DOI: 10.1093/nar/gkaa086

Abstract

The determination of distances between specific points in nucleic acids is essential to understanding their behaviour at the molecular level. The ability to measure distances of 2-10 nm is particularly important: deformations arising from protein binding commonly fall within this range, but the reliable measurement of such distances for a conformational ensemble remains a significant challenge. Using several techniques, we show that electron paramagnetic resonance (EPR) spectroscopy of oligonucleotides spin-labelled with triazole-appended nitroxides at the 2' position offers a robust and minimally perturbing tool for obtaining such measurements. For two nitroxides, we present results from EPR spectroscopy, X-ray crystal structures of B-form spin-labelled DNA duplexes, molecular dynamics simulations and nuclear magnetic resonance spectroscopy. These four methods are mutually supportive, and pinpoint the locations of the spin labels on the duplexes. In doing so, this work establishes 2'-alkynyl nitroxide spin-labelling as a minimally perturbing method for probing DNA conformation.

Authors with CRIS profile

Frank Beierlein Computer-Chemie-Centrum (CCC)

Involved external institutions

University of Oxford GB United Kingdom (GB) University of Kentucky US United States (USA) (US) University of St Andrews GB United Kingdom (GB)

How to cite

APA:

Hardwick, J.S., Haugland, M.M., El-Sagheer, A.H., Ptchelkine, D., Beierlein, F., Lane, A.N.,... Anderson, E.A. (2020). 2'-Alkynyl spin-labelling is a minimally perturbing tool for DNA structural analysis. Nucleic Acids Research, 48(6), 2830-2840. https://doi.org/10.1093/nar/gkaa086

MLA:

Hardwick, Jack S., et al. "2'-Alkynyl spin-labelling is a minimally perturbing tool for DNA structural analysis." Nucleic Acids Research 48.6 (2020): 2830-2840.

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