New pulsed field gradient NMR experiments for the detection of bound water in proteins

Abstract
New H2O-selective homonuclear and heteronuclear 2D NMR experiments have been designed for the observation of protein hydration (PHOGSY, Protein Hydration Observed by Gradient Spectroscop Y). These experiments utilize selective excitation of the H2O resonance and pulsed field gradients for coherence selection and efficient H2O suppression. The method allows for a rapid and sensitive detection of H2O molecules in labelled and unlabelled proteins. In addition it opens a way to measure the residence time of water bound to proteins. Its application to uniformly 15N-labelled FKBP-12 (FK-506 binding protein) is demonstrated.