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Monitoring GPCR conformation with GFP-inspired dyes.
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.
Afiliación
  • Belousov A; Moscow Institute of Physics and Technology, Dolgoprudny 141701, Russia.
  • Maslov I; Moscow Institute of Physics and Technology, Dolgoprudny 141701, Russia.
  • Orekhov P; Dynamic Bioimaging Lab, Advanced Optical Microscopy Centre, Biomedical Research Institute, Agoralaan C (BIOMED), Hasselt University, 3590 Diepenbeek, Belgium.
  • Khorn P; Laboratory for Photochemistry and Spectroscopy, Division for Molecular Imaging and Photonics, Department of Chemistry, KU Leuven, 3001 Leuven, Belgium.
  • Kuzmichev P; Faculty of Biology, Shenzhen MSU-BIT University, Shenzhen 518172, China.
  • Baleeva N; Sechenov University, Moscow 119146, Russia.
  • Motov V; Moscow Institute of Physics and Technology, Dolgoprudny 141701, Russia.
  • Bogorodskiy A; Moscow Institute of Physics and Technology, Dolgoprudny 141701, Russia.
  • Krasnova S; Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117997, Russia.
  • Mineev K; Pirogov Russian National Research Medical University, Moscow 117997, Russia.
  • Zinchenko D; Moscow Institute of Physics and Technology, Dolgoprudny 141701, Russia.
  • Zernii E; Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117997, Russia.
  • Ivanovich V; Moscow Institute of Physics and Technology, Dolgoprudny 141701, Russia.
  • Permyakov S; Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117997, Russia.
  • Hofkens J; National Research University Higher School of Economics, Moscow 101000, Russia.
  • Hendrix J; Moscow Institute of Physics and Technology, Dolgoprudny 141701, Russia.
  • Cherezov V; Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117997, Russia.
  • Gensch T; Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino 142290, Russia.
  • Mishin A; Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119992, Russia.
  • Baranov M; Moscow Institute of Physics and Technology, Dolgoprudny 141701, Russia.
  • Mishin A; Institute for Biological Instrumentation, Russian Academy of Sciences, Pushchino 142292, Russia.
  • Borshchevskiy V; Laboratory for Photochemistry and Spectroscopy, Division for Molecular Imaging and Photonics, Department of Chemistry, KU Leuven, 3001 Leuven, Belgium.
iScience ; 27(8): 110466, 2024 Aug 16.
Article en En | MEDLINE | ID: mdl-39156645
ABSTRACT
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: IScience Año: 2024 Tipo del documento: Article País de afiliación: Rusia Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: IScience Año: 2024 Tipo del documento: Article País de afiliación: Rusia Pais de publicación: Estados Unidos