title: Measurement of Ca²⁺-ATPase activity (in PMCA and SERCA1).
abstract: Ca(2+)-ATP pumps (those on the plasma membrane; PMCA and sarcoplasmic reticulum; SERCA1) have an important role to play in the regulation of intracellular calcium concentrations. In this chapter, three preparations, two membranes and a purified enzyme, best suited for studies of Ca(2+)-ATPase activity are described. The two selected membranes are the human red blood cell (RBC) ghosts, a representative of plasma membranes (PM), and the rabbit skeletal muscle SR, an intracellular membrane. In this protocol, Pi released during the ATPase reaction is subsequently measured colorimetrically as a complex of molybdovanadate. The method is simple (one-step), fast, sensitive, and reliable.
authors.affiliation: Department of Cardiovascular Sciences, Clinical Division of Anaesthesia Critical, Care and Pain Management, University Hospitals of Leicester NHS Trust, Leicester, Leicestershire, UK. DGL3@le.ac.uk
authors.forename: Danuta
authors.initials: D
authors.lastname: Kosk-Kosicka
mesh_terms: D000818:Animals; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D053499:Plasma Membrane Calcium-Transporting ATPases / Q000378:metabolism*; D011817:Rabbits; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism*
chemical_list: D053499:Plasma Membrane Calcium-Transporting ATPases; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D000252:Calcium-Transporting ATPases