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

¹H NMR of Labile Protons: Deuterium (²H) Substitution00:48

¹H NMR of Labile Protons: Deuterium (²H) Substitution

1.3K
This lesson illustrates the role of deuterium substitution in simplifying the NMR spectrum of compounds comprising labile protons. One method employed is the use of deuterium. Amongst the three isotopes of hydrogen, deuterium (2H) has a nucleus composed of one proton and one neutron. When the D2O solvent is added to a pure dry ethanol solution, its labile proton is substituted with deuterium.
1.3K
¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)01:20

¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)

1.6K
When proton-coupled carbon-13 spectra are simplified by a broadband proton decoupling technique, structural information about the coupled protons is lost. Distortionless enhancement by polarization transfer (DEPT) is a technique that provides information on the number of hydrogens attached to each carbon in a molecule. While the DEPT experiment utilizes complex pulse sequences, the pulse delay and flip angle are specifically manipulated. The resulting signals have different phases depending on...
1.6K
Double Resonance Techniques: Overview01:12

Double Resonance Techniques: Overview

680
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.
Spin decoupling is usually achieved by...
680
Hess's Law03:40

Hess's Law

54.7K
There are two ways to determine the amount of heat involved in a chemical change: measure it experimentally, or calculate it from other experimentally determined enthalpy changes. Some reactions are difficult, if not impossible, to investigate and make accurate measurements for experimentally. And even when a reaction is not hard to perform or measure, it is convenient to be able to determine the heat involved in a reaction without having to perform an experiment.
54.7K
¹³C NMR: ¹H–¹³C Decoupling01:04

¹³C NMR: ¹H–¹³C Decoupling

1.7K
The probability of having two carbon-13 atoms next to each other is negligible because of the low natural abundance of carbon-13. Consequently, peak splitting due to carbon-carbon spin-spin coupling is not observed in spectra. However, protons up to three sigma bonds away split the carbon signal according to the n+1 rule, resulting in complicated spectra.
A broadband decoupling technique is used to simplify these complex, sometimes overlapping, signals. Broadband decoupling relies on a...
1.7K
¹H NMR: Complex Splitting01:13

¹H NMR: Complex Splitting

1.8K
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...
1.8K

You might also read

Related Articles

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

Sort by
Same author

Functional roles for FEN1 phosphate steering residues in multi-step substrate verification prior to reaction.

The Journal of biological chemistry·2026
Same author

ANARCII enables alignment-free antigen receptor numbering using a generalised language model.

Communications biology·2026
Same author

iNOS modulates inflammatory responses in an NO-independent manner through direct interaction with IRG1 in mitochondria.

Nature metabolism·2026
Same author

Ginkgo Datapoints Antibody Developability Competition outcomes: limited model performance and a call for data standardization.

mAbs·2026
Same author

LICHEN enables light-chain immunoglobulin sequence generation conditioned on the heavy chain and experimental needs.

Communications biology·2026
Same author

Characterising nanobody developability to improve therapeutic design using the Therapeutic Nanobody Profiler.

Communications biology·2026

Related Experiment Video

Updated: Jan 11, 2026

Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics
09:18

Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics

Published on: April 17, 2017

10.2K

Inferring residue level hydrogen deuterium exchange with ReX.

Oliver M Crook1,2, Nathan Gittens3, Chun-Wa Chung3

  • 1Department of Chemistry, Dorothy Crowfoot Hodgkin Building, University of Oxford, Oxford, UK. oliver.crook@chem.ox.ac.uk.

Communications Chemistry
|November 10, 2025
PubMed
Summary

A new method, ReX, enhances Hydrogen-Deuterium Exchange Mass-Spectrometry (HDX-MS) to provide residue-level protein dynamics. This improves understanding of protein structure and function, aiding drug discovery and mode of action analysis.

More Related Videos

A Hydrogen-Deuterium Exchange Mass Spectrometry HDX-MS Platform for Investigating Peptide Biosynthetic Enzymes
11:32

A Hydrogen-Deuterium Exchange Mass Spectrometry HDX-MS Platform for Investigating Peptide Biosynthetic Enzymes

Published on: May 4, 2020

8.7K
Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry
11:37

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry

Published on: November 29, 2013

18.9K

Related Experiment Videos

Last Updated: Jan 11, 2026

Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics
09:18

Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics

Published on: April 17, 2017

10.2K
A Hydrogen-Deuterium Exchange Mass Spectrometry HDX-MS Platform for Investigating Peptide Biosynthetic Enzymes
11:32

A Hydrogen-Deuterium Exchange Mass Spectrometry HDX-MS Platform for Investigating Peptide Biosynthetic Enzymes

Published on: May 4, 2020

8.7K
Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry
11:37

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry

Published on: November 29, 2013

18.9K

Area of Science:

  • Biochemistry and Structural Biology
  • Computational Biology
  • Analytical Chemistry

Background:

  • Hydrogen-Deuterium Exchange Mass-Spectrometry (HDX-MS) is vital for studying protein dynamics in solution.
  • Current bottom-up HDX-MS methods using peptides limit residue-level resolution and complicate data interpretation.
  • There is a need for enhanced analytical approaches to improve the resolution and interpretability of HDX-MS data.

Purpose of the Study:

  • To introduce ReX, a novel computational method for inferring residue-level deuterium uptake patterns from HDX-MS data.
  • To leverage peptide overlaps, temporal data, and sequence correlations for enhanced resolution.
  • To provide a framework for statistical significance, differential HDX confidence, and uncertainty estimation in protein dynamics.

Main Methods:

  • Developed ReX, a method treating HDX-MS as a multiple change-point problem.
  • Employed a Bayesian non-parametric framework for model fitting and inference.
  • Validated ReX using a three-way proteolytic digestion experiment and benchmarked against existing methods.

Main Results:

  • ReX demonstrates superior performance in predicting unseen HDX data compared to existing methods.
  • The method provides global and local resolution metrics, aligning HDX-MS with other structural biology reporting standards.
  • Applied ReX to analyze differential flexibility in BRD4 and conformational variations in LXRα induced by small molecules.

Conclusions:

  • ReX significantly enhances the resolution and interpretability of HDX-MS data at the residue level.
  • The method accurately characterizes ligand-induced conformational changes, revealing distinct HDX signatures.
  • ReX offers a powerful tool for detailed protein dynamics analysis, informing drug discovery and mode of action studies.