Enantiomeric two-armed lanthanide-binding tags for complementary effects in paramagnetic NMR spectroscopy

Area of Science:

• Biophysical chemistry
• Structural biology
• Nuclear Magnetic Resonance (NMR) spectroscopy

Background:

• Lanthanide-binding tags are essential tools for protein structure determination using NMR.
• Paramagnetic effects from lanthanides provide long-range distance and orientational restraints.
• Developing novel tags with distinct properties is crucial for enhancing structural studies.

Purpose of the Study:

• To develop and characterize novel two-armed lanthanide-binding tags.
• To investigate the paramagnetic effects induced by these tags on protein NMR spectra.
• To enable the measurement of complementary paramagnetic restraints for advanced structural analysis.

Main Methods:

• Synthesis of enantiomeric pairs of rigid, two-armed, cyclen-based lanthanide-binding tags.
• Site-specific attachment of the tags to proteins.
• Acquisition and analysis of NMR spectra to observe paramagnetic effects.
• Utilizing paramagnetic restraints for protein structural modeling.

Main Results:

• The two-armed tags induce significant, long-range paramagnetic effects.
• Enantiomeric tags produce markedly different spectral effects from the same tagging site.
• Complementary restraints derived from both tags enhance structural study capabilities.

Conclusions:

• The developed tags are effective for inducing measurable paramagnetic effects in protein NMR.
• The enantiomeric nature of the tags allows for the collection of complementary structural information.
• These tags represent a valuable advancement for high-resolution protein structure determination.