Assessing multiple conformations of lanthanide binding tags for proteins using a sensitive ¹⁹F-reporter

Area of Science:

• Analytical Chemistry
• Biophysical Chemistry
• Structural Biology

Background:

• Quantifying isomeric species in metal complexes in solution presents significant analytical challenges.
• Lanthanide binding tags are crucial for protein NMR analysis, but their isomeric composition and dynamics are often difficult to assess.
• Developing robust methods for characterizing these tags is essential for advancing protein structure determination.

Purpose of the Study:

• To investigate the utility of fluorine-19 NMR (19F NMR) spectroscopy for quantifying isomeric species of lanthanide binding tags.
• To determine the dynamic properties of these lanthanide binding tags in solution using 19F NMR.
• To assess the efficiency of 19F NMR as a screening tool for paramagnetic tags used in protein NMR.

Main Methods:

• Utilized 19F NMR spectroscopy to analyze samples containing lanthanide binding tags.
• Applied spectral analysis to determine the relative abundance of different isomeric species.
• Monitored changes in NMR signals to probe the dynamic properties of the tags.

Main Results:

• Successfully quantified the isomeric species of lanthanide binding tags in solution.
• 19F NMR provided insights into the dynamic behavior of the investigated tags.
• The results demonstrated a clear correlation between 19F NMR signals and tag characteristics.

Conclusions:

• Fluorine-19 NMR is an effective and efficient method for quantifying isomeric species of lanthanide binding tags.
• 19F NMR serves as a valuable reporter for assessing and screening paramagnetic tags for protein NMR applications.
• This approach facilitates the selection of optimal tags for enhanced protein NMR studies.