De novo design of a fusion protein tool for GPCR research

• 成果类型: Article
• 署名作者: Gao, Kaixuan; Zhang, Xin; Nie, Jia; Meng, Hengyu; Zhang, Weishe; Tian, Boxue; Liu, Xiangyu
• 署名单位: Tsinghua University; Tsinghua University; Central South University
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-12884
• DOI: 10.1073/pnas.2422360122
• 发表日期: 2025-07-22
• 关键词: discovery; m1; platform

摘要

G protein-coupled receptors (GPCRs) play pivotal roles in cellular signaling and represent prominent drug targets. Structural elucidation of GPCRs is crucial for drug discovery efforts. However, structural studies of GPCRs remain challenging, particularly for inactive-state structures, which often require extensive protein engineering. Here, we present a de novo design strategy termed click fusion for generating fusion proteins to facilitate GPCR structural studies. Our method involves the rational design of structurally stable protein domains rigidly linked to GPCRs. The resulting fusion protein enhances the thermostability of the target GPCR and aids in determining GPCR structures via cryoelectron microscopy (cryo-EM). We further demonstrate that the designed fusion protein can be transferred among structurally similar GPCRs with minor adjustments to the linker region. Our study introduces a promising approach for facilitating GPCR structural studies and advancing drug discovery efforts.