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

High-Resolution Mass Spectrometry (HRMS)01:15

High-Resolution Mass Spectrometry (HRMS)

1.2K
The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For...
1.2K
Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

701
Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
701
2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)01:19

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

623
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...
623
Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

881
Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and signal-to-noise ratio for the analyte. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.
Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called collision-induced...
881
2D NMR: Overview of Heteronuclear Correlation Techniques01:18

2D NMR: Overview of Heteronuclear Correlation Techniques

149
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...
149
¹H NMR: Complex Splitting01:13

¹H NMR: Complex Splitting

1.2K
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...
1.2K

You might also read

Related Articles

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

Sort by
Same author

Small Molecule Stabilization of Diverse Amyloidogenic Immunoglobulin Light Chains Revealed by Hydrogen-Deuterium Exchange Mass Spectrometry.

Journal of molecular biology·2026
Same author

Magnesium as a conformational gatekeeper of KRAS: Structural dynamics and therapeutic implications.

Protein science : a publication of the Protein Society·2026
Same author

Structures of DnaA domain I reveal a dimer conserved across Actinomycetes.

Nucleic acids research·2026
Same author

Low-Temperature HILIC Provides Enhanced Separations and Stability for LC-MS-Based Metabolomics.

Journal of proteome research·2026
Same author

Ultraslow conformational dynamics and catch bond formation of a bacterial adhesin revealed by a single-domain variant of FimH.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

More than an attachment module: covalent inhibitor warheads influence BTK dynamics and function.

bioRxiv : the preprint server for biology·2026

Related Experiment Video

Updated: Jun 1, 2025

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

7.9K

Rigorous Analysis of Multimodal HDX-MS Spectra.

Lisa M Tuttle1, Ellie I James2, Florian Georgescauld3

  • 1Department of Biochemistry, University of Washington, Seattle, Washington 98195, United States.

Journal of the American Society for Mass Spectrometry
|January 21, 2025
PubMed
Summary

Hydrogen/deuterium exchange mass spectrometry (HDX-MS) can reveal protein conformational states. New statistical methods and software (HX-Express) accurately analyze multimodal spectra, aiding in the interpretation of complex protein dynamics.

Keywords:
EX1H/D exchangeHDX-MSbimodaldeconvolutiondeuteriummultimodal

More Related Videos

Author Spotlight: Unveiling the Potential of VSFG Microscopy in Studying Mesoscopically Heterogeneous Self-Assembled Structures
08:49

Author Spotlight: Unveiling the Potential of VSFG Microscopy in Studying Mesoscopically Heterogeneous Self-Assembled Structures

Published on: December 1, 2023

1.3K
Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry
11:37

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry

Published on: November 29, 2013

18.3K

Related Experiment Videos

Last Updated: Jun 1, 2025

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

7.9K
Author Spotlight: Unveiling the Potential of VSFG Microscopy in Studying Mesoscopically Heterogeneous Self-Assembled Structures
08:49

Author Spotlight: Unveiling the Potential of VSFG Microscopy in Studying Mesoscopically Heterogeneous Self-Assembled Structures

Published on: December 1, 2023

1.3K
Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry
11:37

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry

Published on: November 29, 2013

18.3K

Area of Science:

  • Biochemistry and Biophysics
  • Analytical Chemistry
  • Structural Biology

Background:

  • Hydrogen/deuterium exchange coupled to mass spectrometry (HDX-MS) is a powerful technique for studying protein dynamics.
  • HDX-MS can detect multiple protein conformational states, often observed as multimodal isotopic envelopes in mass spectra.
  • Established statistical methods for accurately analyzing these multimodal spectra are lacking.

Purpose of the Study:

  • To develop and validate a statistical approach for the accurate detection and deconvolution of multimodal isotopic distributions in HDX-MS data.
  • To provide a robust computational tool for interpreting complex HDX-MS spectra arising from mixed protein populations or complex kinetic events.

Main Methods:

  • An unrestrained binomial distribution fitting approach was developed to analyze isotopic distributions.
  • Statistical tests were implemented to assess the significance of detected subpopulations.
  • The algorithms were integrated into the updated HX-Express software and validated using known peptide mixtures.

Main Results:

  • The developed approach accurately detects and deconvolutes multimodal isotopic envelopes in HDX-MS spectra.
  • Validation using known peptide mixtures demonstrated the reliability of the method.
  • The updated HX-Express software provides a user-friendly tool for analyzing complex HDX-MS data.

Conclusions:

  • This study presents a statistically sound method for analyzing multimodal HDX-MS spectra.
  • The enhanced HX-Express software facilitates the interpretation of protein conformational heterogeneity and complex exchange kinetics.
  • This readily accessible tool will advance the application of HDX-MS in structural biology and biophysics.