A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: Computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and: In situ affinity constant in cells
Class B G protein-coupled receptors (GPCRs) are involved in a variety of human pathophysiological states. These groups of membrane receptors are less studied than class A GPCRs due to the lack of structural information, delayed small molecule drug discovery, and scarce fluorescence detection tools a...
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2018
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_14639076_v20_n46_p29212_Szalai http://hdl.handle.net/20.500.12110/paper_14639076_v20_n46_p29212_Szalai |
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paper:paper_14639076_v20_n46_p29212_Szalai2023-06-08T16:16:33Z A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: Computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and: In situ affinity constant in cells biological marker corticotropin releasing factor receptor corticotropin releasing factor receptor 1 fluorescent dye antagonists and inhibitors chemical structure chemistry fluorescence imaging fluorescence microscopy human molecular docking nanotechnology synthesis Biomarkers Fluorescent Dyes Humans Microscopy, Fluorescence Molecular Docking Simulation Molecular Structure Nanotechnology Optical Imaging Receptors, Corticotropin-Releasing Hormone Class B G protein-coupled receptors (GPCRs) are involved in a variety of human pathophysiological states. These groups of membrane receptors are less studied than class A GPCRs due to the lack of structural information, delayed small molecule drug discovery, and scarce fluorescence detection tools available. The class B corticotropin-releasing hormone type 1 receptor (CRHR1) is a key player in the stress response whose dysregulation is critically involved in stress-related disorders: psychiatric conditions (i.e. depression, anxiety, and addictions), neuroendocrinological alterations, and neurodegenerative diseases. Here, we present a strategy to label GPCRs with a small fluorescent antagonist that permits the observation of the receptor in live cells through stochastic optical reconstruction microscopy (STORM) with 23 nm resolution. The marker, an aza-BODIPY derivative, was designed based on computational docking studies, then synthesized, and finally tested in biological cells. Experiments on hippocampal neurons demonstrate antagonist effects in similar concentrations as the well-established antagonist CP-376395. A quantitative analysis of two color STORM images enabled the determination of the binding affinity of the new marker in the cellular environment. This journal is © the Owner Societies. 2018 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_14639076_v20_n46_p29212_Szalai http://hdl.handle.net/20.500.12110/paper_14639076_v20_n46_p29212_Szalai |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
biological marker corticotropin releasing factor receptor corticotropin releasing factor receptor 1 fluorescent dye antagonists and inhibitors chemical structure chemistry fluorescence imaging fluorescence microscopy human molecular docking nanotechnology synthesis Biomarkers Fluorescent Dyes Humans Microscopy, Fluorescence Molecular Docking Simulation Molecular Structure Nanotechnology Optical Imaging Receptors, Corticotropin-Releasing Hormone |
| spellingShingle |
biological marker corticotropin releasing factor receptor corticotropin releasing factor receptor 1 fluorescent dye antagonists and inhibitors chemical structure chemistry fluorescence imaging fluorescence microscopy human molecular docking nanotechnology synthesis Biomarkers Fluorescent Dyes Humans Microscopy, Fluorescence Molecular Docking Simulation Molecular Structure Nanotechnology Optical Imaging Receptors, Corticotropin-Releasing Hormone A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: Computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and: In situ affinity constant in cells |
| topic_facet |
biological marker corticotropin releasing factor receptor corticotropin releasing factor receptor 1 fluorescent dye antagonists and inhibitors chemical structure chemistry fluorescence imaging fluorescence microscopy human molecular docking nanotechnology synthesis Biomarkers Fluorescent Dyes Humans Microscopy, Fluorescence Molecular Docking Simulation Molecular Structure Nanotechnology Optical Imaging Receptors, Corticotropin-Releasing Hormone |
| description |
Class B G protein-coupled receptors (GPCRs) are involved in a variety of human pathophysiological states. These groups of membrane receptors are less studied than class A GPCRs due to the lack of structural information, delayed small molecule drug discovery, and scarce fluorescence detection tools available. The class B corticotropin-releasing hormone type 1 receptor (CRHR1) is a key player in the stress response whose dysregulation is critically involved in stress-related disorders: psychiatric conditions (i.e. depression, anxiety, and addictions), neuroendocrinological alterations, and neurodegenerative diseases. Here, we present a strategy to label GPCRs with a small fluorescent antagonist that permits the observation of the receptor in live cells through stochastic optical reconstruction microscopy (STORM) with 23 nm resolution. The marker, an aza-BODIPY derivative, was designed based on computational docking studies, then synthesized, and finally tested in biological cells. Experiments on hippocampal neurons demonstrate antagonist effects in similar concentrations as the well-established antagonist CP-376395. A quantitative analysis of two color STORM images enabled the determination of the binding affinity of the new marker in the cellular environment. This journal is © the Owner Societies. |
| title |
A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: Computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and: In situ affinity constant in cells |
| title_short |
A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: Computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and: In situ affinity constant in cells |
| title_full |
A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: Computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and: In situ affinity constant in cells |
| title_fullStr |
A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: Computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and: In situ affinity constant in cells |
| title_full_unstemmed |
A fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: Computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and: In situ affinity constant in cells |
| title_sort |
fluorescence nanoscopy marker for corticotropin-releasing hormone type 1 receptor: computer design, synthesis, signaling effects, super-resolved fluorescence imaging, and: in situ affinity constant in cells |
| publishDate |
2018 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_14639076_v20_n46_p29212_Szalai http://hdl.handle.net/20.500.12110/paper_14639076_v20_n46_p29212_Szalai |
| _version_ |
1768542332294004736 |