title: Enzymatic activity of pig heart mitochondrial malate dehydrogenase monomolecular films by surface exchange technique.
abstract: A technique for studying the catalytic activity of enzymes spread as a film at an air-water interface, by exchanging the subphase under the film to remove unspread enzyme molecules, was developed, and its effectiveness was studied using surface-spread mitochondrial malate dehydrogenase. Mitochondrial malate dehydrogenase formed stable films which gave reproducible pi-A curves. The enzyme activity was measured by the oxidation rate of reduced nicotinamide adenine dinucleotide (NADH) in the presence of the substrate oxalacetic acid. Oxalacetic acid and NADH were injected into the subphase. The catalytic activity of the enzyme was dependent on the surface pressure of the film. The maximum catalytic activity was observed at a surface pressure of 4.4 dynes/cm. The activity was higher at intermediate surface pressures than at very low or very high surface pressures. A high bulk catalytic activity was observed in the unstable region, i.e., at a high degree of compression, of the film. The catalytic activity of the surface-spread enzyme was only a fraction of an equivalent amount of enzyme in solution.
mesh_terms: D000818:Animals; D066298:In Vitro Techniques; D008291:Malate Dehydrogenase / Q000378:metabolism*; D008567:Membranes, Artificial; D008931:Mitochondria, Muscle / Q000201:enzymology*; D009206:Myocardium / Q000201:enzymology* / Q000648:ultrastructure; D011487:Protein Conformation; D013056:Spectrophotometry, Ultraviolet; D013499:Surface Properties; D013500:Surface Tension; D013552:Swine; D013997:Time Factors
chemical_list: D008567:Membranes, Artificial; D008291:Malate Dehydrogenase