Work

Recombinant apoaequorin acting as a pseudo-luciferase reports micromolar changes in the endoplasmic reticulum free Ca2+ of intact cells

Year: 1996

Type: article

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.

Source: Biochemical Journal

DOI: 10.1042/bj3180383

Authors: Jonathan M. KENDALL, Michael N BADMINTON, Graciela B. SALA-NEWBY, Anthony K CAMPBELL, ChristopherM REMBOLD

Institutions: Cardiff University, University of Wales, University of Virginia Medical Center, University of Virginia

Language: English

Topic: bioluminescence and chemiluminescence research

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: bronze

Funders: Wellcome Trust, Medical Research Council, National Heart, Lung, and Blood Institute

Award: G100044611