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

Mostrar todas las versiones(2)

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...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Szalai, A.M., Armando, N.G., Barabas, F.M., Stefani, F.D., Giordano, L., Bari, S.E., Cavasotto, C.N., Silberstein, S., Aramendía, P.F.
Formato: JOUR
Materias:
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
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_14639076_v20_n46_p29212_Szalai
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id todo:paper_14639076_v20_n46_p29212_Szalai
record_format dspace
spelling todo:paper_14639076_v20_n46_p29212_Szalai2023-10-03T16:16:58Z 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 Szalai, A.M. Armando, N.G. Barabas, F.M. Stefani, F.D. Giordano, L. Bari, S.E. Cavasotto, C.N. Silberstein, S. Aramendía, P.F. 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. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar 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
Szalai, A.M.
Armando, N.G.
Barabas, F.M.
Stefani, F.D.
Giordano, L.
Bari, S.E.
Cavasotto, C.N.
Silberstein, S.
Aramendía, P.F.
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.
format JOUR
author Szalai, A.M.
Armando, N.G.
Barabas, F.M.
Stefani, F.D.
Giordano, L.
Bari, S.E.
Cavasotto, C.N.
Silberstein, S.
Aramendía, P.F.
author_facet Szalai, A.M.
Armando, N.G.
Barabas, F.M.
Stefani, F.D.
Giordano, L.
Bari, S.E.
Cavasotto, C.N.
Silberstein, S.
Aramendía, P.F.
author_sort Szalai, A.M.
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
url http://hdl.handle.net/20.500.12110/paper_14639076_v20_n46_p29212_Szalai
work_keys_str_mv AT szalaiam afluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT armandong afluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT barabasfm afluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT stefanifd afluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT giordanol afluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT barise afluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT cavasottocn afluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT silbersteins afluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT aramendiapf afluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT szalaiam fluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT armandong fluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT barabasfm fluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT stefanifd fluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT giordanol fluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT barise fluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT cavasottocn fluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT silbersteins fluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
AT aramendiapf fluorescencenanoscopymarkerforcorticotropinreleasinghormonetype1receptorcomputerdesignsynthesissignalingeffectssuperresolvedfluorescenceimagingandinsituaffinityconstantincells
_version_ 1807322837564784640

Ejemplares similares