Department of Chemistry, Kookmin University, 861-1 Jeongneung-dong, Seongbuk-gu, Seoul, 136-702, Republic of Korea.
New Drug Development Center, Osong Medical Innovation Foundation, Osong Sengmyung-Ro 123, Osong-eup, Heungdeok-gu, Cheongju-si, Chungbuk, Republic of Korea.
Sci Rep. 2017 Aug 8;7(1):7577. doi: 10.1038/s41598-017-08096-6.
G-protein coupled receptors (GPCRs) play indispensable physiological roles in cell proliferation, differentiation, and migration; therefore, identifying the mechanisms by which ligands bind to GPCRs is crucial for developing GPCR-targeting pharmaceutics and for understanding critical biological functions. Although some structural information is available regarding the interactions between GPCRs and their small molecule ligands, knowledge of how GPCRs interact with their corresponding macromolecule ligands, such as peptides and proteins, remains elusive. In this study, we have developed a novel strategy to investigate the precise ligand recognition mechanisms involved in the interaction of endothelin receptor type A (ET) with its ligand, endothelin-1 (ET-1); we call this method "directed degeneration" method. Through flow cytometric screening of a randomized ET library, statistical analysis of the identified sequences, and biochemical studies, the ligand interaction map was successfully obtained.
G 蛋白偶联受体（GPCRs）在细胞增殖、分化和迁移中发挥着不可或缺的生理作用；因此，确定配体与 GPCR 结合的机制对于开发针对 GPCR 的药物以及理解关键的生物学功能至关重要。尽管已经获得了一些关于 GPCR 与其小分子配体之间相互作用的结构信息，但对于 GPCR 如何与其相应的大分子配体（如肽和蛋白质）相互作用的知识仍然难以捉摸。在这项研究中，我们开发了一种新的策略来研究内皮素受体 A（ET）与其配体内皮素-1（ET-1）相互作用中涉及的精确配体识别机制；我们称之为“定向退化”方法。通过流式细胞术筛选随机 ET 文库、对鉴定出的序列进行统计分析以及生化研究，成功获得了配体相互作用图谱。
