Non-radioactive binding assay for bradykinin and angiotensin receptors.
Methods Cell Biol
; 149: 77-85, 2019.
Article
em En
| MEDLINE
| ID: mdl-30616828
ABSTRACT
G protein-coupled receptors (GPCRs) are the largest family of membrane protein playing an important role in cellular signal transduction. GPCRs interact with different molecules acting as ligands capable to trigger responses on signaling pathway. Those molecules present specific binding profiles in which, usually, are determined by methods based on radioactive labeled ligands. Here we present an alternative method based on time-resolved fluorescent labeled ligand, specific customized for angiotensin II receptors (AGTR1 and AGTR2) and kinin receptors (BDKRB1 and BDKRB2) wherein, their natural ligands were labeled with the lanthanide europium to generate the Eu3+-N1-DTT-ligands (AngII, BK, and DBK). Competitive binding profile is determined with a fixed concentration of labeled ligand competing with variable concentrations (10-5 to 10-12) of unlabeled ligand. This method is capable to determine binding profiles with comparable results with traditional one and present a reliable alternative to radioactive based methods usage.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Bioensaio
/
Bradicinina
/
Receptores de Angiotensina
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Animals
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article