Min Hlaing, Sekar Giridhar, Hilty Christian
Chemistry Department, Texas A&M University, 3255 TAMU, College Station, TX 77843 (USA).
Biochemistry Department, Texas A&M University, 2128 TAMU, College Station, TX 77843 (USA).
ChemMedChem. 2015 Sep;10(9):1559-63. doi: 10.1002/cmdc.201500241. Epub 2015 Aug 4.
Nuclear magnetic resonance (NMR) spectroscopy is a valuable technique for ligand screening, because it exhibits high specificity toward chemical structure and interactions. Dissolution dynamic nuclear polarization (DNP) is a recent advance in NMR methodology that enables the creation of non-equilibrium spin states, which can dramatically increase NMR sensitivity. Here, the transfer of such spin polarization from hyperpolarized ligand to protein is observed. Mixing hyperpolarized benzamidine with the serine protease trypsin, a "fingerprint" of enhanced protein signals is observed, which shows a different intensity profile than the equilibrium NMR spectrum of the protein, but coincides closely to the frequency profile of a saturation transfer difference (STD) NMR experiment. The DNP experiment benefits from hyperpolarization and enables observation of all frequencies in a single, rapid experiment. Based on these merits, it is an interesting alternative to the widely used STD experiment for identification of protein-ligand interactions.
核磁共振（NMR）光谱法是一种用于配体筛选的重要技术，因为它对化学结构和相互作用具有高度特异性。溶解动态核极化（DNP）是NMR方法学的一项最新进展，它能够产生非平衡自旋态，从而可显著提高NMR灵敏度。在此，观察到了这种自旋极化从超极化配体向蛋白质的转移。将超极化的苯甲脒与丝氨酸蛋白酶胰蛋白酶混合后，观察到了增强的蛋白质信号“指纹”，其强度分布与蛋白质的平衡NMR光谱不同，但与饱和转移差异（STD）NMR实验的频率分布密切吻合。DNP实验受益于超极化，能够在单个快速实验中观察所有频率。基于这些优点，它是用于鉴定蛋白质 - 配体相互作用的广泛使用的STD实验的一个有趣替代方法。
