CEITEC-Central European Institute of Technology, Masaryk University, Kamenice 5, 625 00, Brno, Czech Republic.
Laboratoire des biomolécules, LBM, Département de chimie, École normale supérieure, PSL University, Sorbonne Université, CNRS, 75005, Paris, France.
Chemistry. 2018 Sep 12;24(51):13418-13423. doi: 10.1002/chem.201802885. Epub 2018 Aug 17.
Hyperpolarized 2D exchange spectroscopy (HYPEX) to obtain high-resolution nuclear magnetic resonance (NMR) spectra of folded proteins under near-physiological conditions is reported. The technique is based on hyperpolarized water, which is prepared by dissolution dynamic nuclear polarization and mixed in situ in an NMR spectrometer with a protein in a physiological saline buffer at body temperature. Rapid exchange of labile protons with the hyperpolarized solvent, combined with cross-relaxation effects (NOEs), leads to boosted signal intensities for many amide H- N correlations in the protein ubiquitin. As the introduction of hyperpolarization to the target protein is mediated via the solvent, the method is applicable to a broad spectrum of target molecules.
报道了一种在近生理条件下获得折叠蛋白质高分辨率核磁共振（NMR）谱的二维极化交换波谱学（HYPEX）技术。该技术基于溶解动态核极化制备的高极化水，在生理盐缓冲液中与体温下的蛋白质在 NMR 光谱仪中就地混合。易质子与高极化溶剂的快速交换，结合交叉弛豫效应（NOE），导致蛋白质泛素中许多酰胺 H-N 相关的信号强度增强。由于极化向靶蛋白的引入是通过溶剂介导的，因此该方法适用于广泛的靶分子。
