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Nuclear Overhauser Enhancement (NOE)01:06

Nuclear Overhauser Enhancement (NOE)

Irradiation of a spin-active nucleus causes an increase or decrease in the signal intensity of neighboring nuclei that are not necessarily chemically bonded or involved in J-coupling. This phenomenon, called the nuclear Overhauser enhancement (NOE), results from through-space interactions between the nuclear spins. The NOE effect decreases with increasing internuclear distance and is generally not observed beyond 4 angstroms. In NOE, dipole-dipole interactions between neighboring spin-active...
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Insensitive Nuclei Enhanced by Polarization Transfer (INEPT)

Insensitive Nuclei Enhanced by Polarization Transfer (INEPT) is an advanced Nuclear Magnetic Resonance (NMR) technique specifically designed to detect and enhance the signals of low-abundance nuclei, such as carbon-13 and nitrogen-15, in small molecules. The fundamental principle behind INEPT is the transfer of polarization from a more abundant and highly polarizable nucleus, typically hydrogen-1, to the low-abundance nucleus of interest. This process effectively boosts the NMR signal of the...
Double Resonance Techniques: Overview01:12

Double Resonance Techniques: Overview

Double resonance techniques in Nuclear Magnetic Resonance (NMR) spectroscopy involve the simultaneous application of two different frequencies or radiofrequency pulses to manipulate and observe two distinct nuclear spins. One important application of double resonance is spin decoupling, which selectively suppresses coupling with one type of nucleus while observing the NMR signal from another nucleus, simplifying the spectrum and enhancing resolution.
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2D NMR: Overview of Homonuclear Correlation Techniques01:16

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Atomic Nuclei: Types of Nuclear Relaxation

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NMR Spectrometers: Resolution and Error Correction

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Related Experiment Video

Updated: Jul 1, 2026

Site Directed Spin Labeling and EPR Spectroscopic Studies of Pentameric Ligand-Gated Ion Channels
11:19

Site Directed Spin Labeling and EPR Spectroscopic Studies of Pentameric Ligand-Gated Ion Channels

Published on: July 4, 2016

Nitroxide spin labeled RNA for long range distance measurements by EPR-PELDOR.

O Frolow1, B Endeward, O Schiemann

  • 1Institut für Organische Chemie und Chemische Biologie (OCCB), Goethe-Universität Frankfurt am Main, Max-von-Laue Str. 7, D-60438 Frankfurt am Main, Germany.

Nucleic Acids Symposium Series (2004)
|September 9, 2008
PubMed
Summary

Site-specific incorporation of nitroxide spin labels enables long-range distance measurements in RNA. Pulsed Electron Double Resonance (PELDOR) successfully determined RNA folds up to 6 nm.

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Published on: July 4, 2016

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Practical Aspects of Sample Preparation and Setup of 1H R1&#961; Relaxation Dispersion Experiments of RNA
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Published on: July 9, 2021

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biophysics

Background:

  • Determining RNA structure is crucial for understanding its biological functions.
  • Long-range distance measurements provide insights into RNA folding and conformation.

Purpose of the Study:

  • To develop a method for site-specific incorporation of nitroxide spin labels into RNA.
  • To apply these labels for long-range distance measurements using Pulsed Electron Double Resonance (PELDOR).

Main Methods:

  • Site-specific incorporation of nitroxide spin labels at uracil (U), cytosine (C), and adenine (A) bases using "on column synthesis".
  • Utilizing Pulsed Electron Double Resonance (PELDOR) spectroscopy for distance measurements.

Main Results:

  • Successful site-specific labeling of RNA bases (U, C, A).
  • PELDOR measurements successfully determined distances in the range of 2-6 nm for several RNA molecules.
  • Demonstrated the feasibility of using this method to probe RNA structure.

Conclusions:

  • Site-specific spin labeling combined with PELDOR is a powerful technique for RNA structural studies.
  • This method allows for the determination of long-range distances, aiding in the elucidation of RNA folds.
  • The developed "on column synthesis" approach is effective for preparing labeled RNA for biophysical analysis.