Structure and organization of full-length epidermal growth factor receptor in extracellular vesicles by cryo-electron tomography

• 成果类型: Article
• 署名作者: Gonzalez-Magaldi, Monica; Gullapalli, Anuradha; Papoulas, Ophelia; Liu, Chang; Leung, Adelaide Y. -H.; Guo, Luqiang; Brilot, Axel F.; Marcotte, Edward M.; Ke, Zunlong; Leahy, Daniel J.
• 署名单位: University of Texas System; University of Texas Austin; University of Texas System; University of Texas Austin; University of Texas System; University of Texas Austin
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-14773
• DOI: 10.1073/pnas.2424678122
• 发表日期: 2025-06-10
• 关键词: crystal-structure; kinase domain; activation; membrane; egfr; expression; affinity; distinct; complex; binding

摘要

We report here transport of full-length epidermal growth factor receptor (EGFR), Insulin Receptor, 7-pass transmembrane receptor Smoothened, and 13-pass Sodium-iodide symporter to extracellular vesicles (EVs) for structural and functional studies. Mass spectrometry confirmed the transported proteins are the most abundant in EV membranes, and the presence of many receptor-interacting proteins in EVs demonstrates their utility for characterizing membrane protein interactomes. Cryo-electron tomography of EGFR-containing EVs reveals that EGFR forms clusters in both the presence and absence of EGF with a similar to 3 nm gap between the inner membrane and cytoplasmic density. EGFR extracellular region (ECR) dimers do not form regular arrays in these clusters. Subtomogram averaging of the 150 kDa EGF-bound EGFR ECR dimer yielded a 15 angstrom map into which the crystal structure of the ligand-bound EGFR ECR dimer fits well. These findings refine our understanding of EGFR activation, clustering, and signaling and establish EVs as a versatile platform for structural and functional characterization of human membrane proteins in cell-derived membranes.