Purification and characterization of a lysophospholipase from human amnionic membranes.
We have identified the presence of a lysophospholipase in human placental tissues and have purified this enzyme from the amnion. The specific activity was highest in the amnion and decreased across adjacent tissues. The purification involved the use of DEAE-Sephadex, phenyl-Sepharose, hydroxylapatite, and sulfylpropyl Sephadex chromatography. The activity of the purified enzyme toward palmitoyl lysophosphatidylcholine is 2.5 mumol min-1 mg-1 and the pH optimum is 7.0. The enzyme is not inhibited by EDTA and does not appear to have a metal ion requirement. The enzyme may be of membrane origin; the purified enzyme requires the presence of detergent during storage. The effects of substrate composition and physical state on enzymatic activity were explored. The enzyme was not active toward mono-, di-, or triglycerides, nor toward diacyl phospholipid. The enzyme was active toward myristoyl and palmitoyl lysophosphatidylcholine at concentrations where these substrates spontaneously form micelles or where Triton X-100 was used to induce co-micellization of the substrate at low concentrations with detergent. A role for this enzyme in processing the lysophospholipid product of phospholipase A action must be considered in evaluating arachidonic acid production in human fetal membranes and placental tissue, particularly during the initiation of labor.
D000650:Amnion / Q000201:enzymology*; D002823:Chorion / Q000201:enzymology; D003656:Decidua / Q000201:enzymology; D003902:Detergents / Q000494:pharmacology; D005260:Female; D006801:Humans; D007700:Kinetics; D008245:Lysophospholipase / Q000302:isolation & purification* / Q000378:metabolism; D017830:Octoxynol; D010740:Phospholipases / Q000302:isolation & purification*; D010920:Placenta / Q000201:enzymology; D011092:Polyethylene Glycols / Q000494:pharmacology; D011247:Pregnancy; D013379:Substrate Specificity; D014018:Tissue Distribution
D003902:Detergents; D011092:Polyethylene Glycols; D017830:Octoxynol; D010740:Phospholipases; D008245:Lysophospholipase