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

Proton (¹H) NMR: Chemical Shift01:07

Proton (¹H) NMR: Chemical Shift

3.1K
Organic molecules primarily contain carbon and hydrogen atoms. While all the hydrogen isotopes are NMR-active, protium or hydrogen-1 is the most abundant. It has a significant energy separation between its nuclear spin states due to its large gyromagnetic ratio. As per Boltzmann's distribution, an increase in the energy separation implies a greater excess population of nuclei available for excitation, resulting in a strong NMR absorption signal.
Absorption signals of all the protium nuclei...
3.1K
¹H NMR: Complex Splitting01:13

¹H NMR: Complex Splitting

1.7K
A proton M that is coupled to a proton X results in doublet signals for M. However, NMR-active nuclei can be simultaneously coupled to more than one nonequivalent nucleus. When M is coupled to a second proton A, such as in styrene oxide, each peak in the doublet is split into another doublet.
Splitting diagrams or splitting tree diagrams are routinely used to depict such complex couplings. While drawing splitting diagrams, the splitting with the larger coupling constant is usually applied...
1.7K
¹H NMR of Conformationally Flexible Molecules: Temporal Resolution00:52

¹H NMR of Conformationally Flexible Molecules: Temporal Resolution

1.2K
At room temperature, the chair conformer of cyclohexane undergoes rapid ring flipping between two equivalent chair conformers at a rate of approximately 105 times per second. These two chair conformers are in equilibrium. The rapid ring flipping results in the interconversion of the axial proton to an equatorial proton and an equatorial to the axial proton. Such interconversions are too rapid and cannot be detected on the NMR timescale. Hence, the NMR spectrometer cannot distinguish between the...
1.2K
¹H NMR Signal Integration: Overview00:58

¹H NMR Signal Integration: Overview

3.2K
The intensity of a signal, which can be represented by the area under the peak, depends on the number of protons contributing to that signal. The area under each peak is shown as a vertical line called an integral, with the integral value listed under it, as seen in the proton NMR spectrum of benzyl acetate. Each integral value is divided by the smallest integral value to obtain the ratio of the number of protons producing each signal. The ratio reveals the relative number of protons and not...
3.2K
NMR Spectroscopy Of Amines01:19

NMR Spectroscopy Of Amines

10.8K
In proton NMR spectroscopy, primary amines and secondary amines showcase their N–H protons as a broad signal in the chemical shift range between δ 0.5 and 5 ppm. The exact position in this range depends on several factors, including sample concentration, hydrogen bonding, and the type of solvent used. Since amine protons undergo fast proton exchange in solution, the protons are labile and therefore do not participate in any splitting with adjacent protons. Thus, the observed peak is...
10.8K
NMR and Mass Spectroscopy of Carboxylic Acids01:30

NMR and Mass Spectroscopy of Carboxylic Acids

5.1K
In ¹H NMR spectroscopy, acidic protons (–COOH) of carboxylic acids are highly deshielded and absorb far downfield, at around 9–12 ppm. The chemical shift value depends on the concentration and solvent used.
While α protons of carboxylic acids absorb at 2–2.5 ppm, β protons absorb further upfield.
Carboxylic acids are easily identified by dissolving them in deuterium oxide, which results in a rapid exchange of the acidic protons with deuterium. This leads to the...
5.1K

You might also read

Related Articles

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

Sort by
Same author

Bromodomain dimers: A case study of BRD4 and family-wide AlphaFold predictions.

Structural dynamics (Melville, N.Y.)·2026
Same author

The NMR Exchange Format (NEF): Specification and Applications.

bioRxiv : the preprint server for biology·2026
Same author

Characterization of flexible RNA binding by tandem RNA recognition motifs through integrative ensemble modelling.

Nucleic acids research·2026
Same author

A fast and efficient strategy for the NMR assignment of Fab methyl groups.

Journal of biomolecular NMR·2026
Same author

<sup>1</sup>H, <sup>13</sup>C, and <sup>15</sup>N resonance assignments and solution structure of the CID domain of SR-related- and CTD-associated factor 8 (SCAF8).

Biomolecular NMR assignments·2025
Same author

Super-resolution triple-resonance NMR spectroscopy for the sequential assignment of proteins.

Science advances·2025
Same journal

Sub1 contributes to heart failure with preserved ejection fraction driven by aging in mice.

Nature communications·2026
Same journal

The BRCA1-A complex restricts replication fork reversal-dependent DNA repair in ATM deficient cells.

Nature communications·2026
Same journal

Signaling downstream of tumor-stroma interaction regulates mucinous colorectal adenocarcinoma apicobasal polarity.

Nature communications·2026
Same journal

Click-polymerized polyenamine membranes for efficient lithium extraction.

Nature communications·2026
Same journal

Joint trajectories of brain atrophy, white matter hyperintensities and cognition quantify brain maintenance.

Nature communications·2026
Same journal

Proton shuttling at electrochemical interfaces under alkaline hydrogen evolution.

Nature communications·2026
See all related articles

Related Experiment Video

Updated: Jan 4, 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

15.9K

Automatic structure-based NMR methyl resonance assignment in large proteins.

Iva Pritišanac1, Julia M Würz1, T Reid Alderson2

  • 1Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance, Goethe University Frankfurt am Main, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany.

Nature Communications
|October 31, 2019
PubMed
Summary
This summary is machine-generated.

Automated MethylFLYA efficiently assigns methyl groups in proteins using nuclear Overhauser effect spectroscopy (NOESY) data. This method significantly improves methyl-based NMR applicability by overcoming resonance assignment challenges.

More Related Videos

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR
14:44

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR

Published on: December 16, 2013

10.0K
Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins
12:47

Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins

Published on: December 27, 2016

19.3K

Related Experiment Videos

Last Updated: Jan 4, 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

15.9K
Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR
14:44

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR

Published on: December 16, 2013

10.0K
Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins
12:47

Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins

Published on: December 27, 2016

19.3K

Area of Science:

  • Biochemistry
  • Structural Biology
  • Nuclear Magnetic Resonance (NMR) Spectroscopy

Background:

  • Isotopically labeled methyl groups serve as NMR probes in large, deuterated proteins.
  • Resonance assignment of these methyl groups is a significant bottleneck for methyl-based NMR.
  • Efficient assignment methods are crucial for broader application in structural biology.

Purpose of the Study:

  • To develop and present an automated method, MethylFLYA, for assigning methyl groups in proteins.
  • To assess the performance and efficiency of MethylFLYA compared to existing algorithms.

Main Methods:

  • MethylFLYA utilizes methyl-methyl nuclear Overhauser effect spectroscopy (NOESY) peak lists for assignment.
  • The method was applied to five proteins ranging from 28-358 kDa, totaling 708 labeled methyl groups.
  • Performance was evaluated based on assignment confidence, accuracy, and runtime.

Main Results:

  • MethylFLYA confidently assigned 488 out of 612 methyl groups with NOESY data (80%).
  • The method demonstrated a high agreement with reference assignments (459/488) and a low error rate (1%).
  • MethylFLYA showed superior performance over alternative algorithms with modest runtimes (0.4-1.2 hours).

Conclusions:

  • MethylFLYA provides an automated and efficient solution for methyl group resonance assignment in NMR.
  • The method enhances the applicability of methyl-based NMR for studying large proteins.
  • MethylFLYA is versatile, handling various labeling patterns and NMR data types.