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. 2025 Jun 10;122(23):e2424678122.
doi: 10.1073/pnas.2424678122. Epub 2025 Jun 2.

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

Monica Gonzalez-Magaldi  1 , Anuradha Gullapalli  1 , Ophelia Papoulas  1 , Chang Liu  1 , Adelaide Y-H Leung  1 , Luqiang Guo  1 , Axel F Brilot  2 , Edward M Marcotte  1 , Zunlong Ke  1   3 , Daniel J Leahy  1
Affiliations

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

Monica Gonzalez-Magaldi et al. Proc Natl Acad Sci U S A. .

Abstract

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 ~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 Å 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.

Keywords: EGFR; cryo-ET; extracellular vesicles; mass spectrometry; subtomogram averaging.

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Conflict of interest statement

Competing interests statement:The authors declare no competing interest.

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