Detection of dynamic substrate binding using MRI

Abstract

Magnetic Resonance Imaging (MRI) is rarely used for molecular binding studies and never without synthetic metallic labels. We designed an MRI approach that can specifically detect the binding of natural substrates (i.e. no chemical labels). To accomplish such detection of substrate-target interaction only, we exploit (i) the narrow resonance of aliphatic protons in free substrate for selective radio-frequency (RF) labeling and, (ii) the process of immobilisation upon binding to a solid-like target for fast magnetic transfer of this label over protons in the target backbone. This cascade of events is ultimately detected with MRI using magnetic interaction between target and water protons. We prove this principle using caffeine as a substrate in vitro and then apply it in vivo in the mouse brain. The combined effects of continuous labeling (label pumping), dynamic reversible binding, and water detection was found to enhance the detection sensitivity by about two to three orders of magnitude.