Products Cited in Publications Worldwide Biomolecular interactions studied by low-field NMR using SABRE hyperpolarization

Biomolecular interactions studied by low-field NMR using SABRE hyperpolarization

Pham, Pierce; Hilty, Christian

DOI: PMID:

Abstract

We demonstrate that low-field NMR provides a means for measuring biomacromol. interactions without requiring a superconducting, or even a permanent magnet. A small mol., 5-fluoropyridine-3-carboximidamide, is designed to be a specific ligand for the trypsin protein, while containing a fluorine atom as a nuclear spin hyperpolarizable label. With hyperpolarization by the parahydrogen based signal amplification by the reversible exchange method, fluorine NMR signals are detectable in the measurement field of 0.85 mT of an electromagnet, at a concentration of less than 100μM. As a weak ligand for the protein, the hyperpolarized mol. can serve as a reporter for measuring the binding of other ligands of interest, illustrated by the determination of the dissociation constant KD of benzamidine from changes in the observed R2 relaxation rates. A signal enhancement of more than 106 compared to Boltzmann polarization at the measurement field indicates that this experiment is not feasible without prepolarization. The extended magnetic field range for the measurement of biomol. interactions under near physiol. conditions, with a protein concentration on the order of 10μM or less, provides a new option for screening of ligand binding, measurement of protein-protein interactions, and measurement of mol. dynamics.

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5-Fluoronicotinonitrile

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