A general method to quantify ligand-driven oligomerization from fluorescence-based images

• Published in: Nature methods Vol. 16; no. 6; pp. 493 - 496
• Main Authors: Stoneman, Michael R., Biener, Gabriel, Ward, Richard J., Pediani, John D., Badu, Dammar, Eis, Annie, Popa, Ionel, Milligan, Graeme, Raicu, Valerică
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.06.2019; Nature Publishing Group
• Subjects: 631/1647/328/1978; 631/1647/328/2057; 631/1647/527/1819; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Brief Communication; Drug delivery; Drug screening; Epidermal growth factor; ErbB Receptors - chemistry; ErbB Receptors - metabolism; Fluorescence; Growth factors; High-throughput screening; Humans; Image Processing, Computer-Assisted - methods; Life Sciences; Ligands; Membrane proteins; Methods; Microscopy, Confocal; Monomers; Oligomerization; Oligomers; Protein Binding; Protein interaction; Protein Interaction Domains and Motifs; Protein Multimerization; Protein-protein interactions; Proteins; Proteomics; Receptors, G-Protein-Coupled - metabolism; Receptors, Gastrointestinal Hormone - metabolism; Secretin; Signal Transduction; Spectrometry; Spectrometry, Fluorescence; Variation
• ISSN: 1548-7091; 1548-7105; 1548-7105
• DOI: 10.1038/s41592-019-0408-9

Here, we introduce fluorescence intensity fluctuation spectrometry for determining the identity, abundance and stability of protein oligomers. This approach was tested on monomers and oligomers of known sizes and was used to uncover the oligomeric states of the epidermal growth factor receptor and the secretin receptor in the presence and absence of their agonist ligands. This method is fast and is scalable for high-throughput screening of drugs targeting protein–protein interactions. Fluorescence intensity fluctuation spectrometry provides a rapid and accurate measurement of the identity, abundance and stability of protein oligomers.