title: A Microplate-Based Bioluminescence Assay of Mitochondrial Calcium Uptake.
abstract: Mitochondrial Ca2+homeostasis is crucial for regulating vital functions such as respiration or apoptosis. Targeted aequorins are excellent probes to measure subcellular Ca2+. Ca2+concentration in mitochondria ([Ca2+]M) is low at rest (about 10-7M) and can increase to the micromolar or even approach the millimolar range, upon cell activation. Here we describe a new quantitative luminescent protocol to directly measure mitochondrial Ca2+uptake, optimized for high throughput. The sensitivity of the method allows detection of changes in either the capacity or the affinity of mitochondrial Ca2+transport.
authors.affiliation: Instituto de Biología y Genética Molecular (IBGM), Universidad de Valladolid y Consejo Superior de Investigaciones Científicas (CSIC), c/ Sanz y Forés 3, 47003, Valladolid, Spain. talonso@ibgm.uva.es.
authors.forename: María Teresa
authors.initials: MT
authors.lastname: Alonso
authors.affiliation: Instituto de Biología y Genética Molecular (IBGM), Universidad de Valladolid y Consejo Superior de Investigaciones Científicas (CSIC), c/ Sanz y Forés 3, 47003, Valladolid, Spain.
authors.forename: Paloma
authors.initials: P
authors.lastname: Navas-Navarro
authors.affiliation: Instituto de Biología y Genética Molecular (IBGM), Universidad de Valladolid y Consejo Superior de Investigaciones Científicas (CSIC), c/ Sanz y Forés 3, 47003, Valladolid, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: García-Sancho
mesh_terms: D000331:Aequorin / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods*; D008928:Mitochondria / Q000378:metabolism*; D013223:Statistics as Topic
chemical_list: D015220:Calcium Channels; C034703:mitochondrial calcium uniporter; D000331:Aequorin; D002118:Calcium
keywords: Aequorin; Calcium uniporter; Plate-reader; Screening