Monitoring protein conformational changes using fluorescent nanoantennas

• Published in: Nature methods Vol. 19; no. 1; pp. 71 - 80
• Main Authors: Harroun, Scott G., Lauzon, Dominic, Ebert, Maximilian C. C. J. C., Desrosiers, Arnaud, Wang, Xiaomeng, Vallée-Bélisle, Alexis
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.01.2022; Nature Publishing Group
• Subjects: 631/1647/2196; 631/45/607; 631/45/612; 631/61/350/59; 631/92/56; Alkaline phosphatase; Alkaline Phosphatase - chemistry; Alkaline Phosphatase - metabolism; Aniline Compounds - chemistry; Antibodies; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biotechnology; Biotin - chemistry; DNA, Single-Stranded - chemistry; Dyes; Dynamic structural analysis; Fluorescence; Fluorescent Dyes - chemistry; High-throughput screening; Life Sciences; Molecular Docking Simulation; Molecular Dynamics Simulation; Monitoring; Nanoantennas; Nanostructures - chemistry; Organophosphorus Compounds - chemistry; Protein Conformation; Protein G; Protein interaction; Proteins; Proteins - chemistry; Proteomics; Screening; Spectrophotometry, Ultraviolet; Substrates
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/s41592-021-01355-5

Understanding the relationship between protein structural dynamics and function is crucial for both basic research and biotechnology. However, methods for studying the fast dynamics of structural changes are limited. Here, we introduce fluorescent nanoantennas as a spectroscopic technique to sense and report protein conformational changes through noncovalent dye-protein interactions. Using experiments and molecular simulations, we detect and characterize five distinct conformational states of intestinal alkaline phosphatase, including the transient enzyme–substrate complex. We also explored the universality of the nanoantenna strategy with another model protein, Protein G and its interaction with antibodies, and demonstrated a rapid screening strategy to identify efficient nanoantennas. These versatile nanoantennas can be used with diverse dyes to monitor small and large conformational changes, suggesting that they could be used to characterize diverse protein movements or in high-throughput screening applications. Fluorescent nanoantennas represent a versatile detection strategy for monitoring fast, large- and small-scale protein dynamics.