Video Experimental Relacionado

Updated: Sep 18, 2025

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Published on: September 13, 2013

Un antagonista del receptor acoplado a proteínas G con fotoswitch etiquetado ilumina la cinética de enlace de

Lars C P Binkhorst¹, Ivana Josimovic¹, Bas De Boer¹

¹Division of Medicinal Chemistry, Amsterdam Institute of Molecular and Life Sciences, Faculty of Science, Vrije Universiteit Amsterdam, De Boelelaan 1108, 1081 HZ Amsterdam, The Netherlands.

Journal of the American Chemical Society

|June 26, 2025

Resumen

Los investigadores desarrollaron el primer ligando fotointercambiable radiomarcado, [3H]VUF26063, para estudiar la cinética de unión dependiente de la luz. Esta herramienta permite el control en tiempo real de la actividad del receptor de histamina H3 (H3R), avanzando la fotofarmacología.

Área de la Ciencia:

Química medicinal
La bioquímica
Farmacología

Sus antecedentes:

La fotofarmacología utiliza ligandos fotorresponsivos para controlar la señalización de proteínas.
Existen investigaciones limitadas sobre la cinética de unión ligando-proteína de los ligandos con capacidad de conmutación fotográfica.
El control espacio-temporal de las proteínas objetivo es crucial en el descubrimiento de fármacos.

Objetivo del estudio:

Descubrir y caracterizar el primer ligando fotointercambiable radiomarcado para evaluar la cinética de unión dependiente de la luz.
Para investigar la cinética de unión inducida por la luz en tiempo real de un ligando fotoswitchable dirigido al receptor de histamina H3 (H3R).

Principales métodos:

Síntesis de un compuesto a base de arilazopirazol (3f) dirigido al H3R.
Etiquetado radiactivo del compuesto para crear [3H]VUF26063 ([3H]3f).
Evaluación en tiempo real de la cinética de unión ligando-proteína utilizando el ligando radiomarcado en condiciones de iluminación variables.

Principales resultados:

[3H]VUF26063 fue sintetizado con éxito y caracterizado como un ligando fotoswitchable.
El ligando demostró afinidad subnanomolar en su isómero trans, con una disminución de 50 veces en la afinidad tras la fotoisomerización.
Se logró un control reversible y dependiente de la luz de la cinética de unión de H3R en tiempo real.
Los datos cinéticos sugieren que 3f se isomeriza dentro de la bolsa de unión H3R al iluminarse.

Conclusiones:

El desarrollo de [3H]VUF26063 proporciona una herramienta novedosa para estudiar las interacciones entre ligandos y proteínas fotoswitchables.
Esta investigación aclara la cinética de unión de los ligandos fotointercambiables, con implicaciones para los GPCR y otros objetivos de proteínas.
Los hallazgos allanan el camino para un control espacial-temporal avanzado en el descubrimiento de fármacos y la biología química.

Más Videos Relacionados

Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding

Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding

Published on: June 9, 2017

Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets

Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets

Published on: March 12, 2019

Videos de Conceptos Relacionados

GPCR Desensitization

GPCR Desensitization

G protein-coupled receptor (GPCR) signaling plays a crucial role in cell functioning. GPCR desensitization is an equally essential process. It allows cells to respond to changing environments and regain sensitivity to new stimuli while preventing unnecessary stimulation when no longer needed. Prolonged exposure to stimuli leads to GPCR desensitization. It involves blocking the receptors from binding and activating additional G proteins. This inhibits activation of downstream effectors, thereby...

The Two-State Receptor Model

The Two-State Receptor Model

The two-state receptor model explains a drug's interaction with receptors, such as G protein-coupled receptors and ligand-gated ion channels, to induce or inhibit a biological response. When no natural ligands are present, a receptor exists in an equilibrium of inactive (Ri) and active (Ra) conformations. The inactive form does not produce a response, while the active form generates a basal effect known as constitutive activity.
The binding affinity of a drug determines its interaction with...

Activation and Inactivation of G Proteins

Activation and Inactivation of G Proteins

Heterotrimeric G proteins are guanine nucleotide-binding proteins. As the name suggests, heterotrimeric G proteins are composed of three subunits: alpha, beta, and gamma. They remain GDP-bound or GTP-bound inside the cells and switch between inactive/active states. The Gα subunit possesses the nucleotide-binding pocket that binds guanine nucleotides and switches between GDP or GTP-bound states. In contrast, the Gꞵ and Gγ subunits are always bound together with high...

G-protein Coupled Receptors

G-protein Coupled Receptors

G-protein coupled receptors are ligand binding receptors that indirectly affect changes in the cell. The actual receptor is a single polypeptide that transverses the cell membrane seven times creating intracellular and extracellular loops. The extracellular loops create a ligand specific pocket which binds to neurotransmitters or hormones. The intracellular loops holds onto the G-protein.

G Protein-coupled Receptors

G Protein-coupled Receptors

G Protein-Coupled Receptors or GPCRs are membrane-bound receptors that transiently associate with heterotrimeric G proteins and induce an appropriate response to sensory stimuli such as light, odors, hormones, cytokines, or neurotransmitters.
GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...