Label-Free Analysis with Multiple Parameters Separates G Protein-Coupled Receptor Signaling Pathways.
Anal Chem
; 92(21): 14509-14516, 2020 11 03.
Article
en En
| MEDLINE
| ID: mdl-33054153
ABSTRACT
Real-time label-free techniques are used to profile G protein-coupled receptor (GPCR) signaling pathways in living cells. However, interpreting the label-free signal responses is challenging, and previously reported methods do not reliably separate pathways from each other. In this study, a continuous angular-scanning surface plasmon resonance (SPR) technique is utilized for measuring label-free GPCR signal profiles. We show how the continuous angular-scanning ability, measuring up to nine real-time label-free parameters simultaneously, results in more information-rich label-free signal profiles for different GPCR pathways, providing a more accurate pathway separation. For this, we measured real-time full-angular SPR response curves for Gs, Gq, and Gi signaling pathways in living cells. By selecting two of the most prominent label-free parameters the full SPR curve angular and intensity shifts, we present how this analysis approach can separate each of the three signaling pathways in a straightforward single-step analysis setup, without concurrent use of signal inhibitors or other response modulating compounds.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Transducción de Señal
/
Resonancia por Plasmón de Superficie
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Receptores Acoplados a Proteínas G
Límite:
Animals
/
Humans
Idioma:
En
Año:
2020
Tipo del documento:
Article