| Autor: |
Szalai AM; Centro de Investigaciones en Bionanociencias-'Elizabeth Jares-Erijman' (CIBION), CONICET, Godoy Cruz 2390, 1425 Ciudad de Buenos Aires, Argentina. pedro.aramendia@cibion.conicet.gov.ar., Armando NG, Barabas FM, Stefani FD, Giordano L, Bari SE, Cavasotto CN, Silberstein S, Aramendía PF |
| Jazyk: |
angličtina |
| Zdroj: |
Physical chemistry chemical physics : PCCP [Phys Chem Chem Phys] 2018 Nov 28; Vol. 20 (46), pp. 29212-29220. |
| DOI: |
10.1039/c8cp06196c |
| Abstrakt: |
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. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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