title: Measuring Ca2+-dependent Ca2+-uptake activity in the mouse heart.
abstract: The apparent Ca(2+) affinity of the isoforms of the sarco/endoplasmic reticulum Ca(2+) ATPase SERCA2 is controlled primarily by two proteins, phospholamban (PLB) and sarcolipin (SLN). The rate of ATP-driven Ca(2+) uptake into sarcoplasmic reticulum (SR)-derived vesicles can be monitored by a technique in which the net uptake of (45)Ca(2+) in the form of an intravesicular calcium oxalate precipitate is recorded. Here, we present details of a modification of such a protocol for determining the apparent Ca(2+) affinity of the Ca(2+) pump, and its control by various regulators, in crude homogenates of mouse heart.
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, B3000 Leuven, Belgium.
authors.forename: Tine
authors.initials: T
authors.lastname: Holemans
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, B3000 Leuven, Belgium.
authors.forename: Ilse
authors.initials: I
authors.lastname: Vandecaetsbeek
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, B3000 Leuven, Belgium.
authors.forename: Frank
authors.initials: F
authors.lastname: Wuytack
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, B3000 Leuven, Belgium.
authors.forename: Peter
authors.initials: P
authors.lastname: Vangheluwe
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D045424:Complex Mixtures / Q000378:metabolism; D051379:Mice; D009206:Myocardium / Q000201:enzymology* / Q000378:metabolism*; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism*
chemical_list: D045424:Complex Mixtures; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; C506947:Atp2a2 protein, mouse; D002118:Calcium