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Two-Dimensional (2D) NMR: Overview01:12

Two-Dimensional (2D) NMR: Overview

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The 1D NMR spectrum of large and complex molecules like natural products has complicated splitting patterns and overlapping signals, which can be easily interpreted using 2-dimensional (2D) NMR. Unlike 1D NMR, 2D NMR has two frequency axes that provide the coupling information between the nucleus A and nucleus B in a molecule. The process from which 2D spectra are obtained has four steps.
The first step is the preparation period, during which nucleus A is excited with a radiofrequency pulse....
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2D NMR: Overview of Homonuclear Correlation Techniques01:16

2D NMR: Overview of Homonuclear Correlation Techniques

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Homonuclear correlation spectroscopy (COSY) is a powerful technique used in Nuclear Magnetic Resonance (NMR) spectroscopy to study the correlations between nuclei of the same type within a molecule. It provides information about scalar couplings between adjacent nuclei, which helps determine connectivity and structural information. There are several COSY variants, each with its unique strengths and experimental parameters.
COSY90 is the standard two-dimensional (2D) COSY experiment that...
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NMR Spectrometers: Radiofrequency Pulses and Pulse Sequences01:17

NMR Spectrometers: Radiofrequency Pulses and Pulse Sequences

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A pulse is a short burst of radio waves distributed over a range of frequencies that simultaneously excites all the nuclei in the sample. Upon passing a radio frequency pulse along the x-axis, the nuclei absorb energy corresponding to their Larmor frequencies and achieve resonance. This shifts the net magnetization vector from the z-axis toward the transverse plane. This angle of rotation of the magnetization vector, or the flip angle, is proportional to the duration and intensity of the pulse.
1.8K
NMR Spectrometers: Resolution and Error Correction01:14

NMR Spectrometers: Resolution and Error Correction

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When magnetic nuclei in a sample achieve resonance and undergo relaxation, the signal detected in NMR is an approximately exponential free induction decay. Fourier transform of an exponential decay yields a Lorentzian peak in the frequency domain. Lorentzian peaks in an NMR spectrum are defined by their amplitude, full width at half maximum, and position, where the peak width is governed by the spin-spin relaxation time alone. In real experiments, however, the applied magnetic field is rendered...
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2D NMR: Overview of Heteronuclear Correlation Techniques01:18

2D NMR: Overview of Heteronuclear Correlation Techniques

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Heteronuclear correlation spectroscopy is an analytical technique that investigates the coupling between different types of nuclei, often a proton and an X-nucleus, such as carbon-13 or nitrogen-15. This method is commonly used in nuclear magnetic resonance (NMR) spectroscopy to gain insights into complex chemical compounds' structural and compositional aspects. A typical heteronuclear correlation spectrum displays X-nucleus chemical shifts on one axis and a proton spectrum on the other...
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¹H NMR of Labile Protons: Temporal Resolution01:10

¹H NMR of Labile Protons: Temporal Resolution

1.4K
Protons bonded to heteroatoms such as nitrogen and oxygen exhibit a range of chemical shift values. This is due to the varying degree of hydrogen bonding between the proton and the heteroatom in other molecules. The extent of hydrogen bonding affects the electron density around the proton, thereby giving different chemical shift values for the protons in the proton NMR spectrum.
The –OH proton in alcohols typically appears in the range of δ 2 to 5 ppm but can vary depending on the...
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Related Experiment Video

Updated: May 3, 2026

Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments
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Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments

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Time-resolved multidimensional NMR with non-uniform sampling.

Maxim Mayzel1, Joakim Rosenlöw, Linnéa Isaksson

  • 1The Swedish NMR Centre, University of Gothenburg, Box 465, 40530, Göteborg, Sweden.

Journal of Biomolecular NMR
|January 18, 2014
PubMed
Summary
This summary is machine-generated.

This study introduces a new 3D NMR method for fast, high-resolution kinetic analysis of complex biological reactions. It enables detailed study of protein dynamics and modifications, even in crowded spectral regions.

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Area of Science:

  • Biophysics
  • Biochemistry
  • Structural Biology

Background:

  • Time-resolved experiments require high spectral and temporal resolution.
  • Multidimensional NMR experiments offer high spectral resolution but are limited by lengthy data collection times.
  • Studying dynamic processes in complex biological systems like protein phosphorylation is challenging.

Purpose of the Study:

  • To develop a novel method for high-resolution, time-resolved multidimensional NMR experiments.
  • To overcome the limitations of long data acquisition times in kinetic studies.
  • To enable quantitative site-specific analysis of fast biological processes.

Main Methods:

  • Acquisition of a continuous fast pulsing 3D NMR experiment using non-uniform sampling.
  • Simultaneous processing of the full dataset with multi-dimensional decomposition for enhanced sensitivity and time-resolution.
  • Application to measure deuterium exchange rates in ubiquitin and in vitro phosphorylation kinetics of CD79b.

Main Results:

  • Achieved high sensitivity and time-resolution of a few minutes.
  • Successfully verified the method with simulations and experimental data.
  • Enabled quantitative site-specific kinetic analysis of protein phosphorylation in a crowded spectral region.

Conclusions:

  • Modern NMR data collection and signal processing offer a solution for time-resolved multidimensional experiments.
  • The developed 3D time-resolved NMR approach significantly enhances spectral resolution for kinetic studies.
  • This method is crucial for characterizing complex biological processes, such as protein phosphorylation, with high precision.