title: Monitoring yeast intracellular Ca2+ levels using an in vivo bioluminescence assay.
abstract: This protocol describes the use of the jellyfish Aequorea victoria aequorin protein to measure Ca(2+) levels in living yeast cells. All yeast strains to be analyzed must express the A. victoria apoprotein of the aequorin calcium biosensor, to be reconstituted into fully active aequorin by association with its cofactor, coelenterazine, which cannot be synthesized by yeast itself. The simplest way to achieve reconstitution is to transform yeast cells with a vector driving apoaequorin expression, and then supply commercially available coelenterazine cofactor in the medium. Coelenterazine is a hydrophobic molecule and is able to permeate yeast cells.
authors: Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Milan, Italy;
authors: Renata
authors: R
authors: Tisi
authors: Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Milan, Italy;
authors: Enzo
authors: E
authors: Martegani
authors: Laboratório de Bioquímica e Biotecnologia de Leveduras, Núcleo de Pesquisas em Ciências Biológicas, Escola de Farmácia, Universidade Federal de Ouro Preto, Campus do Morro do Cruzeiro, 35.400-000 Ouro Preto, Minas Gerais, Brazil.
authors: Rogelio L
authors: RL
authors: Brandão
mesh_terms: D002118:Calcium / Q000378:metabolism*; D008163:Luminescent Measurements; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
chemical_list: D002118:Calcium