Work

Quantitative Mapping of Nanoscale EGFR–Grb2 Assemblies by DNA‐PAINT

Year: 2026

Type: article

Abstract: Receptor tyrosine kinase signaling is initiated by extracellular ligand binding, which drives the formation of membrane-protein assemblies that activate intracellular signal transduction. Accurately resolving the molecular composition of these assemblies in situ remains challenging due to their nanoscale dimensions and intrinsic heterogeneity. Here, we introduce a single-molecule super-resolution imaging and analysis workflow designed to resolve and quantitatively characterize individual membrane-protein assembly sites in cells. We apply this approach to the nanoscale organization of the epidermal growth factor receptor (EGFR) and its adaptor protein Grb2 following stimulation with the native ligand epidermal growth factor. As activation progresses, we observe a reduction in EGFR density at the plasma membrane, a progressive accumulation of Grb2 at EGFR assembly sites, and an increase in both dimeric and higher-order oligomeric EGFR. The experimental and analytical framework presented here is broadly applicable to the study of diverse membrane-protein assemblies.

Source: ChemPhysChem

DOI: 10.1002/cphc.202500915

Authors: Alexandra Kaminer, Yunqing Li, Hans‐Dieter Barth, Marina S. Dietz, Mike Heilemann

Institutions: Goethe University Frankfurt, Max Planck Institute of Biophysics

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $4,550

Funder: Deutsche Forschungsgemeinschaft