title: A continuous spectrophotometric method for monitoring phospholipase D-catalyzed reactions of physiological substrates.
abstract: A continuous spectrophotometric method for monitoring phospholipase D-catalyzed hydrolysis of long acyl chain phosphatidylcholines has been formulated at pH 8.0 in a mixed detergent system using the coupling enzymes choline oxidase and peroxidase. Standard curves for phosphatidylcholine determination in both end-point and rate modes are presented and applied to the estimation of that phospholipid in a solubilized human erythrocyte membrane sample. In rate mode the method is suitable for kinetic study of phospholipase D with physiological substrates in micellar form.
authors.affiliation: Department of Chemistry and Biochemistry, University of Windsor, Ont., Canada.
authors.forename: J F
authors.initials: JF
authors.lastname: Takrama
authors.forename: K E
authors.initials: KE
authors.lastname: Taylor
mesh_terms: D002384:Catalysis; D003902:Detergents; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008991:Monitoring, Physiologic / Q000379:methods; D010713:Phosphatidylcholines / Q000378:metabolism*; D010739:Phospholipase D / Q000378:metabolism*; D012995:Solubility; D013053:Spectrophotometry
chemical_list: D003902:Detergents; D010713:Phosphatidylcholines; D010739:Phospholipase D