title: ATPase Activity Measurements by an Enzyme-Coupled Spectrophotometric Assay.
abstract: Enzymatic coupled assays are usually based on the spectrophotometric registration of changes in NADH/NAD(+) or NADPH/NADP(+) absorption at 340 nm accompanying the oxidation/reduction of reactants that by dehydrogenases and other helper enzymes are linked to the activity of the enzymatic reaction under study. The present NADH-ATP-coupled assay for ATPase activity is a seemingly somewhat complicated procedure, but in practice adaptation to performance is easily acquired. It is a more safe and elegant method than colorimetric methods, but not suitable for handling large number of samples, and also presupposes that the activity of the helper enzymes is not severely affected by the chemical environment of the sample in which it is tested.
authors.affiliation: Department of Biomedicine, Aarhus University, Aarhus C, Denmark.|Centre for Membrane Pumps in Cells and Disease, Danish National Research Foundation, Aarhus C, Denmark.
authors.forename: Pankaj
authors.initials: P
authors.lastname: Sehgal
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, Aarhus C, Denmark.|Centre for Membrane Pumps in Cells and Disease, Danish National Research Foundation, Aarhus C, Denmark.
authors.forename: Claus
authors.initials: C
authors.lastname: Olesen
authors.affiliation: Department of Biomedicine, Aarhus University, Aarhus C, Denmark. jvm@biomed.au.dk.|Centre for Membrane Pumps in Cells and Disease, Danish National Research Foundation, Aarhus C, Denmark. jvm@biomed.au.dk.
authors.forename: Jesper V
authors.initials: JV
authors.lastname: Møller
mesh_terms: D000251:Adenosine Triphosphatases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D003124:Colorimetry / Q000379:methods; D057075:Enzyme Assays / Q000379:methods*; D006868:Hydrolysis; D007700:Kinetics; D009243:NAD / Q000737:chemistry; D009249:NADP / Q000737:chemistry; D010084:Oxidation-Reduction; D013053:Spectrophotometry / Q000379:methods*
chemical_list: D009243:NAD; D009249:NADP; D000251:Adenosine Triphosphatases
keywords: ATP hydrolysis; Ca2+ -ATPase; Coupled assay; Enzymatic spectrophotometry; NADH; SERCA