Department of Chemistry and Biology, Ryerson University, Toronto, Ontario, Canada.
Department of Biochemistry and Molecular Biomedicine, Institute of Biomedicine, University of Barcelona, Barcelona, Spain.
FASEB J. 2019 Mar;33(3):3841-3850. doi: 10.1096/fj.201800440RR. Epub 2018 Dec 6.
Equilibrative nucleoside transporters (ENTs) translocate nucleosides and nucleobases across plasma membranes, as well as a variety of anti-cancer, -viral, and -parasite nucleoside analogs. They are also key members of the purinome complex and regulate the protective and anti-inflammatory effects of adenosine. Despite their important role, little is known about the mechanisms involved in their regulation. We conducted membrane yeast 2-hybrid and coimmunoprecipitation studies and identified, for the first time to our knowledge, the existence of protein-protein interactions between human ENT1 and ENT2 (hENT1 and hENT2) proteins in human cells and the formation of hetero- and homo-oligomers at the plasma membrane and the submembrane region. The use of NanoLuc Binary Technology allowed us to analyze changes in the oligomeric status of hENT1 and hENT2 and how they rapidly modify the uptake profile for nucleosides and nucleobases and allow cells to respond promptly to external signals or changes in the extracellular environment. These changes in hENTs oligomerization are triggered by PKC activation and subsequent action of protein phosphatase 1.-Grañe-Boladeras, N., Williams, D., Tarmakova, Z., Stevanovic, K., Villani, L. A., Mehrabi, P., Siu, K. W. M., Pastor-Anglada, M., Coe, I. R. Oligomerization of equilibrative nucleoside transporters: a novel regulatory and functional mechanism involving PKC and PP1.
平衡核苷转运体（ENTs）可将核苷和碱基跨质膜转运，以及各种抗癌、抗病毒和抗寄生虫核苷类似物。它们也是嘌呤体复合物的关键成员，调节腺苷的保护和抗炎作用。尽管它们的作用很重要，但对其调节机制知之甚少。我们进行了膜酵母 2 杂交和共免疫沉淀研究，首次在人类细胞中发现了人类 ENT1 和 ENT2（hENT1 和 hENT2）蛋白之间的蛋白-蛋白相互作用，并在质膜和亚膜区形成了异源和同源寡聚体。NanoLuc Binary Technology 的使用使我们能够分析 hENT1 和 hENT2 的寡聚状态的变化，以及它们如何快速改变核苷和碱基的摄取谱，使细胞能够迅速对外界信号或细胞外环境的变化做出反应。这些 hENTs 寡聚化的变化是由 PKC 激活和随后的蛋白磷酸酶 1 作用触发的。-Grañe-Boladeras，N.，Williams，D.，Tarmakova，Z.，Stevanovic，K.，Villani，L. A.，Mehrabi，P.，Siu，K. W. M.，Pastor-Anglada，M.，Coe，I. R. 平衡核苷转运体的寡聚化：一种涉及 PKC 和 PP1 的新型调节和功能机制。
