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 Conformationally Flexible Molecules: Temporal Resolution00:52

¹H NMR of Conformationally Flexible Molecules: Temporal Resolution

1.3K
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...
1.3K
¹H NMR of Labile Protons: Temporal Resolution01:10

¹H NMR of Labile Protons: Temporal Resolution

1.8K
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 specific...
1.8K
¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)01:20

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

1.8K
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...
1.8K
¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR01:15

¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR

1.8K
The axial and equatorial protons in cyclohexane can be distinguished by performing a variable-temperature NMR experiment. In this process, except for one proton, the remaining eleven protons are replaced by deuterium. The deuterium substitution avoids the possible peak splitting caused by the spin-spin coupling between the adjacent protons. The remaining proton flips between the axial and equatorial positions.
1.8K

You might also read

Related Articles

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

Sort by
Same author

Correction: Dynamic conformational equilibria in the active states of KRAS and NRAS.

RSC chemical biology·2026
Same author

A <sup>13</sup>C<sup>β</sup> CEST experiment with improved sensitivity for the characterization of protein excited states.

Journal of magnetic resonance (San Diego, Calif. : 1997)·2026
Same author

The dynamic and heterogeneous structure of the non-canonical inflammasome.

bioRxiv : the preprint server for biology·2026
Same author

Making invisible excited-state structures of pro-interleukin-18 visible by combining NMR and machine learning.

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

Structural heterogeneity and substrate engagement mechanism of the bacterial proteasome activator Bpa.

Nature communications·2026
Same author

Probing Submillisecond-to-Millisecond Time Scale Conformational Dynamics in High-Molecular-Weight Biomolecules via <sup>15</sup>N Clean Transverse-Relaxation-Optimized Chemical Exchange Saturation Transfer (TROSY-CEST).

The journal of physical chemistry letters·2025

Related Experiment Video

Updated: Feb 27, 2026

Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments
09:25

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

Published on: November 1, 2024

2.9K

Probing slow timescale dynamics in proteins using methyl 1H CEST.

Tairan Yuwen1, Rui Huang1, Lewis E Kay2,3

  • 1Departments of Molecular Genetics, Biochemistry and Chemistry, University of Toronto, Toronto, ON, Canada.

Journal of Biomolecular NMR
|June 26, 2017
PubMed
Summary

A new 1H CEST experiment enables robust analysis of protein conformational exchange by eliminating proton-proton NOE dips. This method accurately extracts methyl proton chemical shifts for rare protein conformers.

Keywords:
13CH3-/13CHD2-methyl labeling1H CESTConformational exchangeMethyl-TROSYms timescale dynamics

More Related Videos

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
Study of Protein Dynamics via Neutron Spin Echo Spectroscopy
08:03

Study of Protein Dynamics via Neutron Spin Echo Spectroscopy

Published on: April 13, 2022

2.6K

Related Experiment Videos

Last Updated: Feb 27, 2026

Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments
09:25

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

Published on: November 1, 2024

2.9K
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
Study of Protein Dynamics via Neutron Spin Echo Spectroscopy
08:03

Study of Protein Dynamics via Neutron Spin Echo Spectroscopy

Published on: April 13, 2022

2.6K

Area of Science:

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

Background:

  • 15N- and 13C-based Chemical Exchange Saturation Transfer (CEST) are established techniques for studying biomolecular conformational exchange.
  • 1H CEST experiments are emerging as a valuable tool but face challenges with proton-proton Nuclear Overhauser Effect (NOE) interference.

Purpose of the Study:

  • To develop and validate a methyl-TROSY 1H CEST experiment for robust analysis of protein conformational dynamics.
  • To enable the extraction of methyl proton chemical shifts for transient or rare protein conformers.

Main Methods:

  • Development of a methyl-TROSY 1H CEST pulse sequence designed to suppress detrimental 1H-1H NOE dips.
  • Optimization of the experiment for proteins labeled with 13CHD2 groups.
  • Application of the developed method to study protein systems exhibiting conformational exchange.

Main Results:

  • The methyl-TROSY 1H CEST experiment effectively eliminates 1H-1H NOE artifacts, allowing for reliable CEST profile analysis.
  • Accurate methyl proton chemical shifts of rare protein conformers were successfully extracted.
  • Demonstrated utility in studying exchanging protein systems.

Conclusions:

  • The presented methyl-TROSY 1H CEST experiment provides a robust method for characterizing protein conformational exchange.
  • This technique complements existing methods like methyl 1H CPMG, offering a powerful tool for structural biology research.