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

Molecular-orientation analysis based on alignment-induced TROSY chemical shift changes.

Shin-ichi Tate1, Hideto Shimahara, Naoko Utsunomiya-Tate

  • 1Department of structural biology, Biomolecular Engineering Research Institute, 6-2-3 Furuedai, Suita, Osaka 565-0874, Japan. tate@beri.or.jp

Journal of Magnetic Resonance (San Diego, Calif. : 1997)
|November 18, 2004
PubMed
Summary
This summary is machine-generated.

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

Time-resolved proteomics reveals five modules of stress granule disassembly and identifies SYNCRIP as a late-phase clearance factor.

The international journal of biochemistry & cell biology·2026
Same author

Valosin-containing protein counteracts ATP-driven dissolution of FUS condensates through its ATPase activity in vitro.

FEBS letters·2026
Same author

Potential roles of aspartic acid D-isomerization in proteins implicated in the pathogenesis of Alzheimer's disease.

Neural regeneration research·2026
Same author

Site-Specific Aspartic Acid d-Isomerization in Tau R2 and R3 Peptide Seeds Attenuates Seed-Induced Fibril Formation of Full-Length Tau.

Biomolecules·2026
Same author

ISRIB Promotes Aggregation of TIA-1 by Modulating Its Phase Separation in Vitro.

Biological & pharmaceutical bulletin·2025
Same author

Controlled Molecular Orders in Layered Multiple Porphyrins.

Journal of the American Chemical Society·2025

This study introduces a novel NMR method using alignment-induced chemical shift changes in transverse relaxation optimized spectroscopy (TROSY) to determine protein alignment tensors. This technique offers a valuable alternative to traditional methods for analyzing weakly aligned proteins.

Area of Science:

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

Background:

  • Determining the alignment tensor of weakly aligned proteins is crucial for structural studies.
  • Traditional methods often rely on residual dipolar couplings (RDCs), which can be challenging for large proteins.
  • Alignment-induced chemical shift changes in NMR provide valuable information about anisotropic spin interactions.

Purpose of the Study:

  • To develop and validate a new NMR technique for determining protein alignment tensors using only alignment-induced 15N TROSY chemical shift changes.
  • To demonstrate the utility of this method as an alternative to RDC-based approaches, particularly for large, weakly aligned proteins.

Main Methods:

  • Utilized alignment-induced 15N TROSY chemical shift changes, which reflect residual dipolar couplings (RDCs) and residual chemical shift anisotropy effects (RCSAs).

Related Experiment Videos

  • Developed a TROSY-based analysis to extract alignment tensor values from these chemical shift changes.
  • Validated the method by comparing alignment angles with RDC-determined values for 15N-labeled ubiquitin.

    • Main Results:

    • The TROSY-based analysis yielded alignment angles consistent with RDC-based methods for ubiquitin (8.6 kDa), with a maximum uncertainty of 5 degrees.
    • The method demonstrated robustness despite variations in literature-reported 15N chemical shift anisotropy (CSA) tensor values, yielding consistent orientation angles.
    • The alignment tensor could be determined even with pre-determined 15N CSA tensor uncertainties.

    Conclusions:

    • Alignment-induced 15N TROSY chemical shift changes can effectively determine a weakly aligned protein's alignment tensor.
    • This TROSY-based approach is a viable and useful alternative to RDC-based methods for determining alignment angles, especially for large proteins.
    • The method provides reliable structural information, overcoming limitations of existing techniques.