Measuring Protein-Ligand Binding by Hyperpolarized Ultrafast NMR

Area of Science:

• Biophysical Chemistry
• Nuclear Magnetic Resonance (NMR) Spectroscopy
• Structural Biology

Background:

• Protein-ligand interactions are crucial in biological processes and drug discovery.
• Traditional methods for studying these interactions can be time-consuming and require significant sample amounts.
• Nuclear relaxation rates, particularly transverse relaxation rates (R2), are sensitive to molecular binding events.

Purpose of the Study:

• To develop and validate an enhanced NMR technique for rapid detection of protein-ligand interactions.
• To leverage ultrafast NMR and dissolution dynamic nuclear polarization (D-DNP) for improved sensitivity and speed.
• To enable simultaneous measurement of multiple relaxation rates for detailed structural insights.

Main Methods:

• Utilized an ultrafast NMR technique correlating chemical shift with transverse relaxation rate (R2).
• Combined the NMR method with dissolution dynamic nuclear polarization (D-DNP) for signal enhancement.
• Acquired R2 values for multiple carbon spins in unlabeled benzylamine within a single scan.

Main Results:

• Achieved thousands-fold signal enhancement using D-DNP, making R2 values observable in a single scan.
• The hyperpolarized ultrafast chemical shift-R2 correlated experiment successfully separated chemical shift encoding and readout phases.
• Demonstrated the ability to overcome spectral resolution limits imposed by the sampling theorem.

Conclusions:

• The developed technique enables rapid and sensitive detection of protein-ligand interactions.
• Simultaneous acquisition of multiple relaxation rates provides valuable structural information.
• This approach holds significant potential for studying the structural properties of protein-ligand complexes.