Recombinant apoaequorin acting as a pseudo-luciferase reports micromolar changes in the endoplasmic reticulum free Ca2+ of intact cells.
Biochem J
; 318 ( Pt 2): 383-7, 1996 Sep 01.
Article
en En
| MEDLINE
| ID: mdl-8809023
ABSTRACT
We describe a novel method to monitor the endoplasmic reticulum (ER) free Ca2+ in intact cells. Continuous perfusion of HeLa cells, expressing ER-targeted apoaequorin, with coelenterazine allowed the apoprotein to act as a pseudo-luciferase capable of reporting free Ca2+ from 0.1-100 microM. In intact HeLa cells, addition of ionomycin increased apoaequorin-generated light by 91%, indicating that resting ER free Ca2+ was approx. 2 microM. Agonist stimulation decreased the ER apoaequorin signal and proportionally increased cytosolic free Ca2+ consistent with agonist-induced release of Ca2+ from the ER.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Apoproteínas
/
Pirazinas
/
Calcio
/
Aequorina
/
Retículo Endoplásmico
/
Imidazoles
/
Luciferasas
Tipo de estudio:
Diagnostic_studies
Límite:
Humans
Idioma:
En
Año:
1996
Tipo del documento:
Article