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

Sensitivity enhancement in (HCA)CONH experiments.

R Folmer1, G Otting

  • 1Structural Chemistry Laboratory, AstraZeneca R&D Mölndal, Sweden.

Journal of Biomolecular NMR
|May 11, 2000
PubMed
Summary

A new method enhances protein resonance experiments by replenishing lost magnetization, improving sensitivity by 1.6-fold for the (HCA)CONH experiment compared to HN(CA)CO.

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

Compact, hydrophilic, lanthanide-binding tags for paramagnetic NMR spectroscopy.

Chemical science·2018
Same author

RIDME distance measurements using Gd(iii) tags with a narrow central transition.

Physical chemistry chemical physics : PCCP·2016
Same author

Protein conformation by EPR spectroscopy using gadolinium tags clicked to genetically encoded p-azido-L-phenylalanine.

Chemical communications (Cambridge, England)·2015
Same author

Selective excitation of intense solvent signals in the presence of radiation damping.

Journal of biomolecular NMR·2012
Same author

Water-protein NOEs: Optimized scheme for selective water excitation.

Journal of biomolecular NMR·2010
Same author

Okadaic acid inhibits relaxant neural transmission in rat gastric fundus in vitro.

Acta physiologica Scandinavica·2002

Area of Science:

  • Structural Biology
  • Nuclear Magnetic Resonance (NMR) Spectroscopy
  • Protein Dynamics

Background:

  • Protein backbone resonance assignment is crucial for understanding protein structure and function.
  • Dipole-CSA cross-correlated relaxation leads to magnetization loss in NMR experiments.
  • Existing techniques face limitations in sensitivity and efficiency.

Purpose of the Study:

  • To introduce a novel sensitivity-enhancement technique for protein NMR experiments.
  • To improve the correlation of protein backbone resonances using 13Calpha-1Halpha groups.
  • To overcome sensitivity losses caused by dipole-CSA cross-correlated relaxation.

Main Methods:

  • Developed a technique to replenish 13Calpha spin magnetization lost via dipole-CSA cross-correlated relaxation.
  • Applied the principle to the (HCA)CONH experiment.
  • Evaluated variations including spin-locking of transverse 13C-1H two-spin coherence and a cross-correlation compensated (CA)CONH experiment.

Main Results:

  • Achieved a 1.6-fold sensitivity enhancement for the (HCA)CONH experiment compared to the standard HN(CA)CO experiment.
  • Demonstrated the effectiveness of magnetization replenishment in improving signal-to-noise ratio.
  • Investigated alternative experimental setups for further optimization.

Conclusions:

  • The proposed sensitivity-enhancement technique significantly improves protein NMR experiment efficiency.
  • This method provides a valuable tool for structural and dynamic studies of proteins.
  • Further development of related experiments can broaden their applicability in structural biology.

Related Experiment Videos