PELDOR Measurements on Nitroxide-Labeled Oligonucleotides

Tobias Hett, Olav Schiemann

Publikation: Beitrag in Buch/Bericht/KonferenzbandKapitelBegutachtung

Abstract

In the past decades, pulsed dipolar electron paramagnetic resonance spectroscopy (PDS) has emerged as a powerful tool in biophysical chemistry to study the structure, dynamics, and function of biomolecules like oligonucleotides and proteins. Structural information is obtained from PDS methods in form of a distribution of distances between spin centers. Such spin centers can either be intrinsically present paramagnetic metal ions and organic radicals or may be attached to the biomolecule by means of site-directed spin labeling. The most common PDS experiment for probing interspin distances in the nanometer range is pulsed electron–electron double resonance (PELDOR or DEER). In the protocol presented here, we provide a step-by-step workflow on how to set up a PELDOR experiment on a commercially available pulsed EPR spectrometer, outline the data analysis, and highlight potential pitfalls. We suggest PELDOR measurements on nitroxide-labeled oligonucleotides to study the structure of either RNA-cleaving DNAzymes in complex with their RNA targets or modified DNAzymes with different functions and targets, in which deoxynucleotides are substituted by nitroxide-labeled nucleotides.

OriginalspracheEnglisch
TitelMethods in Molecular Biology
Herausgeber (Verlag)Humana Press Inc.
Seiten241-274
Seitenumfang34
DOIs
PublikationsstatusVeröffentlicht - 2022
Extern publiziertJa

Publikationsreihe

NameMethods in Molecular Biology
Band2439
ISSN (Print)1064-3745
ISSN (elektronisch)1940-6029

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