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 Experiment Videos

A high sensitivity 3D experiment for measuring Calpha-Halpha residual dipolar coupling constants.

Weidong Hu1, Ziming Zhang, Yuan Chen

  • 1Division of Immunology, Beckman Institute of the City of Hope, Duarte, CA 91010, USA. whu@coh.org

Journal of Magnetic Resonance (San Diego, Calif. : 1997)
|December 4, 2003
PubMed
Summary

A novel nuclear magnetic resonance (NMR) method enhances sensitivity for measuring protein couplings. This improved HA(CA)CONH approach offers accurate scalar and residual dipolar coupling measurements in labeled proteins.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Inhibition of ADSS2-mediated de novo AMP biosynthesis re-sensitizes acute myeloid leukemia to BH3 mimetics.

Nature cancer·2026
Same author

Targeting the METTL1/m7G axis as a therapeutic strategy in myeloid leukemia.

Blood·2026
Same author

International Consensus on Severe Lung Cancer-The Second Edition.

Translational lung cancer research·2026
Same author

Physical parameters estimation for Michelson interferometric fringes based on FFARNet-18.

Optics express·2026
Same author

Comparative <i>In Vitro</i> Analysis of Five Starch Sources: Nutrient Release Patterns and Functional Effects in a Simulated Gastrointestinal Environment.

Foods (Basel, Switzerland)·2026
Same author

Allosteric inhibition of RAN decreases miR-126 biogenesis in endothelial cells and controls acute myeloid leukemia growth.

Communications biology·2026

Area of Science:

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

Background:

  • Measuring Calpha-Halpha scalar and dipolar couplings is crucial for protein structure determination.
  • Previous methods, like the (HA)CA(CO)NH sequence, faced limitations in sensitivity.
  • High-sensitivity techniques are needed for analyzing complex or low-concentration protein samples.

Purpose of the Study:

  • To present a new, sensitivity-enhanced nuclear magnetic resonance (NMR) approach for measuring Calpha-Halpha scalar and dipolar coupling constants.
  • To compare the sensitivity and performance of the new HA(CA)CONH scheme against existing methods.
  • To demonstrate the applicability of the new method on a biologically relevant protein target.

Main Methods:

  • Development and implementation of the HA(CA)CONH NMR sequence for 13C/15N-labeled proteins.

Related Experiment Videos

  • Comparison of sensitivity with the previously established (HA)CA(CO)NH sequence.
  • Application of the new sequence to the C-terminal domain of the human Ku-80 protein.

    • Main Results:

    • The HA(CA)CONH scheme demonstrated significantly higher sensitivity compared to the (HA)CA(CO)NH sequence.
    • Average sensitivity improvement of 40% was observed for both isotropic and anisotropic samples.
    • Sensitivity enhancement was more pronounced in structured protein regions (50-60%) compared to unstructured regions.

    Conclusions:

    • The novel HA(CA)CONH NMR approach provides a substantial sensitivity improvement for measuring Calpha-Halpha couplings.
    • This method allows for accurate and straightforward determination of scalar and residual dipolar coupling constants.
    • The sequence is readily implementable and effective for studying protein structure and dynamics, as shown with the human Ku-80 protein.