title: Effect of diolein on hydrolysis of phosphatidylcholine by phospholipase C from Clostridium perfringens.
abstract: The activity of phospholipase C from Clostridium perfringens on 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) as a monolayer at an air/water interface was examined. With a pure POPC monolayer, sharp cut-off of the enzyme activity was observed on increase in surface pressure. However, this cut-off disappeared on addition of a 0.3 molar fraction of 1,2-dioleoylglycerol (1,2-DO) to the monolayer. An abrupt change in the enzyme activity was observed with molar fractions of between 0.2 and 0.3 1,2-DO in the POPC monolayer at an initial surface pressure of 35 mN/m. For examination of the effect of 1,2-DO on the phospholipase C activity, the quantity of [125I]phospholipase C adsorbed to the surface was determined. The enzyme was found to be adsorbed nonspecifically to all lipid films except that of POPC only. The adsorption of enzyme was not affected by the presence or absence of Ca2+ and Zn2+. The rate constant for enzyme adsorption to a 1,2-DO film was 4.5 times that for its adsorption to a POPC film. The adsorption decreased linearly with increase in the surface concentration of POPC, and increased with increase in the surface concentration of 1,2-DO. These data suggest that 1,2-DO (a reaction product) regulates the interaction of phospholipase C with films containing substrate and may also regulate the enzyme activity.
authors.affiliation: Department of Medical Biochemistry, School of Medicine, Ehime University.
authors.forename: T
authors.initials: T
authors.lastname: Tsujita
authors.forename: H
authors.initials: H
authors.lastname: Okuda
mesh_terms: D003016:Clostridium perfringens / Q000201:enzymology*; D004075:Diglycerides / Q000494:pharmacology*; D006868:Hydrolysis; D010713:Phosphatidylcholines / Q000378:metabolism*; D010738:Type C Phospholipases / Q000378:metabolism*
chemical_list: D004075:Diglycerides; D010713:Phosphatidylcholines; D010738:Type C Phospholipases; C028694:1-palmitoyl-2-oleoylphosphatidylcholine; C013965:diolein