Hydrolysis of supported pyrenephospholipid monolayers by phospholipase A2.
Hydrolysis by pancreatic and snake venom (Crotalus atrox) phospholipase A2 of fluorescent monolayers of pyrene-labelled phosphatidylglycerol on solid support was studied. We used a fluorescence microscope equipped with video camera, video recorder and an image analyzer to monitor changes in fluorescence. Decrease in pyrene excimer emission was evident when pyrene phosphatidylglycerol monolayers transferred onto quartz glass slides (at a surface pressure of 15 mN m-1) were subjected to enzymatic hydrolysis. Snake venom phospholipase A2 could hydrolyze the monolayers almost completely while pancreatic phospholipase A2 could cause only 50% decrease in fluorescence intensity. EDTA totally inhibited the action of both A2 phospholipases. When monolayers were transferred onto solid supports at a surface pressure of 31 mN m-1 C. atrox phospholipase A2 could still exert activity whereas porcine pancreatic phospholipase A2 was inactive.
Department of Medical Chemistry, University of Helsinki, Finland.
T
T
Thuren
K K
KK
Eklund
J A
JA
Virtanen
P K
PK
Kinnunen
D003435:Crotalid Venoms / Q000032:analysis; D006868:Hydrolysis; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence; D010179:Pancreas / Q000201:enzymology; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D010743:Phospholipids / Q000032:analysis*; D011721:Pyrenes / Q000032:analysis*
D003435:Crotalid Venoms; D008567:Membranes, Artificial; D010743:Phospholipids; D011721:Pyrenes; D010741:Phospholipases A; D054467:Phospholipases A2