Section of Organic Chemistry and Biochemistry, Department of Chemistry, University of Ioannina, Greece.
Department of Pharmaceutical Chemistry, College of Health Sciences, University of KwaZulu-Natal (Westville Campus), Durban, South Africa.
FEBS Lett. 2018 Oct;592(20):3367-3379. doi: 10.1002/1873-3468.13250. Epub 2018 Sep 29.
In-cell NMR spectroscopy has emerged as a powerful technique for monitoring biomolecular interactions at an atomic level inside intact cells. However, current methodologies are inadequate at charting intracellular interactions of nonlabeled proteins and require their prior isotopic labeling. Herein, we describe for the first time the monitoring of the quercetin-alanine bioconjugate interaction with the nonlabeled antiapoptotic protein Bcl-2 inside living human cancer cells. STD and Tr-NOESY in-cell NMR methodologies were successfully applied in the investigation of the binding, which was further validated in vitro. In-cell NMR proved a very promising strategy for the real-time probing of the interaction profile of potential drugs with their therapeutic targets in native cellular environments and could, thus, open a new avenue in drug discovery.
细胞内 NMR 光谱学已成为一种强大的技术，可在完整细胞内原子水平监测生物分子相互作用。然而，目前的方法在绘制未标记蛋白质的细胞内相互作用方面还不够完善，需要对其进行同位素标记。在此，我们首次描述了在活的人类癌细胞内监测槲皮素-丙氨酸生物缀合物与非标记抗凋亡蛋白 Bcl-2 的相互作用。STD 和 Tr-NOESY 细胞内 NMR 方法成功应用于结合的研究，进一步在体外进行了验证。细胞内 NMR 为实时探测潜在药物与其治疗靶点在天然细胞环境中的相互作用模式提供了一种很有前途的策略，因此，可能为药物发现开辟新途径。
