Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

2D NMR: Overview of Homonuclear Correlation Techniques01:16

2D NMR: Overview of Homonuclear Correlation Techniques

698
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...
698
2D NMR: Overview of Heteronuclear Correlation Techniques01:18

2D NMR: Overview of Heteronuclear Correlation Techniques

821
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...
821
2D NMR: Homonuclear Correlation Spectroscopy (COSY)01:06

2D NMR: Homonuclear Correlation Spectroscopy (COSY)

2.0K
Homonuclear correlation spectroscopy, or COSY, is a 2-dimensional NMR technique that provides information about coupled protons. Typically, the geminal and vicinal coupling are observed. For example, consider the COSY spectrum of ethyl acetate, where its 1D proton NMR spectrum is plotted along the vertical and horizontal axes with their corresponding chemical shift scale. Three spots on the diagonal corresponding to the three peaks in the 1D proton spectrum are called diagonal peaks. The COSY...
2.0K
NMR Spectrometers: Overview01:20

NMR Spectrometers: Overview

2.3K
NMR spectrometers consist of a strong magnet, a radiofrequency transmitter, and a detector attached to a computer console for recording spectra of samples containing NMR-active nuclei. In first-generation NMR instruments called continuous-wave spectrometers, the resonance frequencies of the nuclei are determined by frequency-sweep or field-sweep methods. The magnetic field strength is fixed and the rf signal is swept in the former, while the radiofrequency signal is fixed and the magnetic field...
2.3K
NMR Spectroscopy: Spin–Spin Coupling01:08

NMR Spectroscopy: Spin–Spin Coupling

3.3K
The spin state of an NMR-active nucleus can have a slight effect on its immediate electronic environment. This effect propagates through the intervening bonds and affects the electronic environments of NMR-active nuclei up to three bonds away; occasionally, even farther. This phenomenon is called spin–spin coupling or J-coupling. Coupling interactions are mutual and result in small changes in the absorption frequencies of both nuclei involved. While nuclei of the same element are involved...
3.3K
2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)01:19

2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)

1.5K
Heteronuclear single-quantum correlation spectroscopy (HSQC) is a 2D NMR technique that reveals one-bond correlations between hydrogen and a heteronucleus. The HSQC experiment is similar to the heteronuclear correlation experiment (HETCOR) but is more sensitive. In the HSQC spectrum, the proton chemical shift is plotted on the horizontal F2 axis, while the 13C chemical shift is plotted on the vertical F1 axis. The corresponding proton and 13C spectra are also shown. The HSQC contour plot does...
1.5K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Probing Substrate-Loaded Carrier Proteins by Nuclear Magnetic Resonance.

Methods in molecular biology (Clifton, N.J.)·2023
Same author

Dynamics and mechanistic interpretations of nonribosomal peptide synthetase cyclization domains.

Current opinion in chemical biology·2022
Same author

High-resolution structures of a siderophore-producing cyclization domain from Yersinia pestis offer a refined proposal of substrate binding.

The Journal of biological chemistry·2022
Same author

<sup>15</sup>N-Detected TROSY NMR experiments to study large disordered proteins in high-field magnets.

Chemical communications (Cambridge, England)·2022
Same author

Global protein dynamics as communication sensors in peptide synthetase domains.

Science advances·2022
Same author

NMR as a readout to monitor and restore the integrity of complex chemoenzymatic reactions.

Journal of magnetic resonance (San Diego, Calif. : 1997)·2022
Same journal

Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Post-Selection Methods for Analyzing mRNA Display Selections and Optimization of Hits.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

High-Performance Computing in Tandem Mass Spectrometry (MS/MS) Peptide Identification.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Engineering and Adapting Disulfide-Containing Proteins to Enable Intracellular Functionality.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

AI-Driven Protein Research: From Prediction to Design.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Methods for the In Vitro Selection of Protein and Peptide Libraries Using mRNA Display.

Methods in molecular biology (Clifton, N.J.)·2026
See all related articles

Related Experiment Video

Updated: Feb 18, 2026

Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy
14:55

Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy

Published on: September 17, 2017

16.1K

Covariance NMR Processing and Analysis for Protein Assignment.

Bradley J Harden1, Dominique P Frueh2

  • 1Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, 701 Hunterian, 725 N Wolfe St., Baltimore, MD, 21205, USA.

Methods in Molecular Biology (Clifton, N.J.)
|November 20, 2017
PubMed
Summary
This summary is machine-generated.

Covariance NMR (Nuclear Magnetic Resonance) offers a robust method for assigning protein resonances by correlating nuclei indirectly. New processing steps significantly reduce artifacts, enabling reliable 4D covariance map analysis for challenging protein assignments.

Keywords:
4D spectraCovarianceNMRPeak listsResonance assignmentSpectral derivative

More Related Videos

Author Spotlight: Unveiling the Structural and Dynamic Aspects of Glycan Molecular Recognition
07:40

Author Spotlight: Unveiling the Structural and Dynamic Aspects of Glycan Molecular Recognition

Published on: May 17, 2024

2.0K
Exploring Protein-Glycan Interactions: Advances in Nuclear Magnetic Resonance
10:07

Exploring Protein-Glycan Interactions: Advances in Nuclear Magnetic Resonance

Published on: August 26, 2025

609

Related Experiment Videos

Last Updated: Feb 18, 2026

Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy
14:55

Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy

Published on: September 17, 2017

16.1K
Author Spotlight: Unveiling the Structural and Dynamic Aspects of Glycan Molecular Recognition
07:40

Author Spotlight: Unveiling the Structural and Dynamic Aspects of Glycan Molecular Recognition

Published on: May 17, 2024

2.0K
Exploring Protein-Glycan Interactions: Advances in Nuclear Magnetic Resonance
10:07

Exploring Protein-Glycan Interactions: Advances in Nuclear Magnetic Resonance

Published on: August 26, 2025

609

Area of Science:

  • Structural Biology
  • Biophysics
  • Nuclear Magnetic Resonance Spectroscopy

Background:

  • Nuclear Magnetic Resonance (NMR) is crucial for determining protein structure and dynamics.
  • Indirect correlation of nuclei is often required for comprehensive NMR resonance assignment.
  • Covariance NMR (COV-NMR) provides a powerful alternative to traditional peak picking for correlating nuclei, but is prone to artifacts.

Purpose of the Study:

  • To present a detailed protocol for applying pre- and postprocessing steps to reduce artifacts in COV-NMR spectra.
  • To enable the calculation and interpretation of 4D COV-NMR spectra for protein resonance assignment.
  • To demonstrate the utility of enhanced COV-NMR for assigning resonances in large, complex proteins.

Main Methods:

  • Development and application of pre- and postprocessing scripts for COV-NMR data.
  • Generation of 4D covariance maps from pairs of 3D NMR spectra.
  • Systematic assignment of backbone and sidechain resonances in two challenging protein samples.

Main Results:

  • Significant reduction in false-positive artifacts in COV-NMR spectra.
  • Successful generation of diverse 4D covariance maps.
  • Effective assignment of NMR signals in large and difficult protein targets.

Conclusions:

  • The enhanced COV-NMR protocol effectively mitigates artifacts, improving spectral reliability.
  • 4D covariance maps are valuable tools for complex protein resonance assignment.
  • COV-NMR, with improved processing, is poised to become integral to NMR signal assignment strategies.