Monitoring GPCR conformation with GFP-inspired dyes

Solvatochromic compounds have emerged as valuable environment-sensitive probes for biological research. Here we used thiol-reactive solvatochromic analogs of the green fluorescent protein (GFP) chromophore to track conformational changes in two proteins, recoverin and the A2A adenosine receptor (A2A...

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Published in:iScience 2024-08, Vol.27 (8), p.110466, Article 110466
Main Authors: Belousov, Anatoliy, Maslov, Ivan, Orekhov, Philipp, Khorn, Polina, Kuzmichev, Pavel, Baleeva, Nadezhda, Motov, Vladislav, Bogorodskiy, Andrey, Krasnova, Svetlana, Mineev, Konstantin, Zinchenko, Dmitry, Zernii, Evgeni, Ivanovich, Valentin, Permyakov, Sergei, Hofkens, Johan, Hendrix, Jelle, Cherezov, Vadim, Gensch, Thomas, Mishin, Alexander, Baranov, Mikhail, Mishin, Alexey, Borshchevskiy, Valentin
Format: Article
Language:English
Subjects:
Biochemistry
Biophysics
Structural biology
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Summary:Solvatochromic compounds have emerged as valuable environment-sensitive probes for biological research. Here we used thiol-reactive solvatochromic analogs of the green fluorescent protein (GFP) chromophore to track conformational changes in two proteins, recoverin and the A2A adenosine receptor (A2AAR). Two dyes showed Ca2+-induced fluorescence changes when attached to recoverin. Our best-performing dye, DyeC, exhibited agonist-induced changes in both intensity and shape of its fluorescence spectrum when attached to A2AAR; none of these effects were observed with other common environment-sensitive dyes. Molecular dynamics simulations showed that activation of the A2AAR led to a more confined and hydrophilic environment for DyeC. Additionally, an allosteric modulator of A2AAR induced distinct fluorescence changes in the DyeC spectrum, indicating a unique receptor conformation. Our study demonstrated that GFP-inspired dyes are effective for detecting structural changes in G protein-coupled receptors (GPCRs), offering advantages such as intensity-based and ratiometric tracking, redshifted fluorescence spectra, and sensitivity to allosteric modulation. [Display omitted] •GFP-inspired dyes can detect structural changes in GPCRs•DyeC responds to changes induced by allosteric modulator and orthosteric agonists•Conformational changes can be tracked ratiometrically and by fluorescence intensity Biochemistry; Structural biology; Biophysics
ISSN:2589-0042
2589-0042
DOI:10.1016/j.isci.2024.110466