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Related Experiment Video

Updated: Jan 30, 2026

Paramagnetic Relaxation Enhancement for Detecting and Characterizing Self-Associations of Intrinsically Disordered Proteins
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Long-Range RNA Structural Information via a Paramagnetically Tagged Reporter Protein.

Madeleine Strickland1, Jonathan Catazaro, Rohith Rajasekaran1

  • 1Laboratory of Structural Biophysics, Biochemistry and Biophysics Center, National Heart, Lung and Blood Institute , National Institutes of Health , Bethesda , Maryland 20892 , United States.

Journal of the American Chemical Society
|January 18, 2019
PubMed
Summary
This summary is machine-generated.

This study introduces a novel method for determining the structure of large RNA molecules using Nuclear Magnetic Resonance (NMR) and paramagnetic labeling. This technique enables atomic-level structural studies of complex RNA.", Enhanced_Abstract=default_api.SeocontentEnhancedAbstract(Area_of_Science=["Structural Biology", "Biophysics", "Molecular Biology"], Background=["Nuclear Magnetic Resonance (NMR) provides structural and dynamical insights for small RNA molecules (up to ~50 nucleotides).", "Studying larger RNA structures with NMR is challenging due to difficulties in establishing global structural features.", "Paramagnetic labeling is effective for proteins but limited for larger RNAs due to challenges in site-specific labeling."], Purpose_of_the_Study=["To develop a strategy for site-specific paramagnetic labeling of large RNA molecules for structural determination using NMR.", "To enable atomic-level structural studies of RNA molecules previously inaccessible to detailed NMR analysis."], Main_Methods=["Modification of RNA loop residues to facilitate binding to a paramagnetically tagged reporter protein (U1A RNA-binding domain).", "Measurement of Lanthanide-induced pseudocontact shifts (PCS) in complexed RNA.", "Validation of the method using a 232-nucleotide RNA and a 36-nucleotide RNA with known structures."], Main_Results=["Demonstrated successful application of lanthanide-induced pseudocontact shifts for a 232-nucleotide RNA bound to tagged U1A derivatives.", "Validated the method by showing agreement between measured NMR values and predicted values for a 36-nucleotide RNA.", "Established a broadly applicable approach for atomic-level study of large RNAs."], Conclusions=["The developed strategy allows for the atomic-level structural investigation of large RNA molecules.", "The ability to insert U1A binding sites into RNA structures makes this method versatile.", "This approach overcomes previous limitations in applying NMR-based paramagnetic labeling to large RNAs."]), Meta_Description=

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Area of Science:

  • Structural Biology
  • Biophysics
  • Molecular Biology

Background:

  • Nuclear Magnetic Resonance (NMR) provides structural and dynamical insights for small RNA molecules (up to ~50 nucleotides).
  • Studying larger RNA structures with NMR is challenging due to difficulties in establishing global structural features.
  • Paramagnetic labeling is effective for proteins but limited for larger RNAs due to challenges in site-specific labeling.

Purpose of the Study:

  • To develop a strategy for site-specific paramagnetic labeling of large RNA molecules for structural determination using NMR.
  • To enable atomic-level structural studies of RNA molecules previously inaccessible to detailed NMR analysis.

Main Methods:

  • Modification of RNA loop residues to facilitate binding to a paramagnetically tagged reporter protein (U1A RNA-binding domain).
  • Measurement of Lanthanide-induced pseudocontact shifts (PCS) in complexed RNA.
  • Validation of the method using a 232-nucleotide RNA and a 36-nucleotide RNA with known structures.

Main Results:

  • Demonstrated successful application of lanthanide-induced pseudocontact shifts for a 232-nucleotide RNA bound to tagged U1A derivatives.
  • Validated the method by showing agreement between measured NMR values and predicted values for a 36-nucleotide RNA.
  • Established a broadly applicable approach for atomic-level study of large RNAs.

Conclusions:

  • The developed strategy allows for the atomic-level structural investigation of large RNA molecules.
  • The ability to insert U1A binding sites into RNA structures makes this method versatile.
  • This approach overcomes previous limitations in applying NMR-based paramagnetic labeling to large RNAs.