Enzymatic assays: optimization of systems by using pyruvate kinase and lactate dehydrogenase as auxiliary enzymes.
A method to optimize enzymatic assays by using pyruvate kinase and lactate dehydrogenase enzymes is presented and applied to mitochondrial ATPase as an example. Optimum amounts of auxiliary enzymes, to obtain either a 99% of the initial rate in a given time (t99) or a given lag period (L), are calculated from their apparent Michaelis constants (Kapp) in the medium used and their prices per enzymatic international unit.
D000818:Animals; D002417:Cattle; D003362:Cost-Benefit Analysis; D007700:Kinetics; D007770:L-Lactate Dehydrogenase; D008433:Mathematics; D008722:Methods; D008928:Mitochondria / Q000201:enzymology; D010086:Oxidative Phosphorylation Coupling Factors / Q000032:analysis; D006180:Proton-Translocating ATPases / Q000032:analysis*; D011770:Pyruvate Kinase
D010086:Oxidative Phosphorylation Coupling Factors; D007770:L-Lactate Dehydrogenase; D011770:Pyruvate Kinase; D006180:Proton-Translocating ATPases