Phillips Margaret, To Janet, Yamazaki Toshio, Nagashima Toshio, Torres Jaume, Pervushin Konstantin
School of Biological Sciences, Nanyang Technological University, Singapore, 637551, Singapore.
RIKEN Centre for Life Science Technologies, Kanagawa, 230-0045, Japan.
J Biomol NMR. 2018 Jun;71(2):91-100. doi: 10.1007/s10858-018-0195-0. Epub 2018 Jun 18.
Aquaporins are integral membrane proteins that facilitate water flow across biological membranes. Their involvement in multiple physiological functions and disease states has prompted intense research to discover water channel activity modulators. However, inhibitors found so far are weak and/or lack specificity. For organic compounds, which lack of high electron-dense atoms, the identification of binding sites is even more difficult. Nuclear magnetic resonance spectroscopy (NMR) requires large amounts of the protein, and expression and purification of mammalian aquaporins in large quantities is a difficult task. However, since aquaporin Z (AqpZ) can be purified and expressed in good quantities and has a high similarity to human AQP1 (~ 40% identity), it can be used as a model for studying the structure and function of human aquaporins. In the present study, we have used solid-state MAS NMR to investigate the binding of a lead compound [1-(4-methylphenyl)1H-pyrrole-2,5-dione] to AqpZ, through mapping of chemical shift perturbations in the presence of the compound.
水通道蛋白是整合膜蛋白，可促进水跨生物膜流动。它们参与多种生理功能和疾病状态，这促使人们进行深入研究以发现水通道活性调节剂。然而，迄今为止发现的抑制剂作用较弱且/或缺乏特异性。对于缺乏高电子密度原子的有机化合物，确定其结合位点更加困难。核磁共振光谱法（NMR）需要大量蛋白质，而大量表达和纯化哺乳动物水通道蛋白是一项艰巨任务。然而，由于水通道蛋白Z（AqpZ）可以大量纯化和表达，并且与人类水通道蛋白1具有高度相似性（约40%的同一性），因此它可作为研究人类水通道蛋白结构和功能的模型。在本研究中，我们使用固态核磁共振波谱法，通过在化合物存在下绘制化学位移扰动图谱，研究了先导化合物[1-(4-甲基苯基)-1H-吡咯-2,5-二酮]与AqpZ的结合。
