Enhancement of Agkistrodon piscivorus piscivorus venom phospholipase A2 activity toward phosphatidylcholine vesicles by lysolecithin and palmitic acid: studies with fluorescent probes of membrane structure.
The activity of phospholipase A2 from snake venom to hydrolyze bilayers of phosphatidylcholines is greatly enhanced by the presence of the hydrolysis products, lysolecithin and fatty acid, in the bilayer. The fluorescence of several probes of membrane structure was used to monitor changes in bilayer physical properties during vesicle hydrolysis. These changes were compared to emission spectra and fluorescence polarization results occurring upon direct addition of lysolecithin and/or fatty acid to the bilayer. The excimer to monomer ratio of 1,3-bis(1-pyrene)propane was insensitive to vesicle hydrolysis, suggesting that changes in the order of the phospholipid chains were not relevant to the effect of the hydrolysis products on phospholipase activity. The fluorescence of 6-propionyl-2-(dimethylamino)-naphthalene (Prodan) suggested that the polarity of the bilayer in the region of the phospholipid head groups increases as the hydrolysis products accumulate in the bilayer. The fluorescence of 6-dodecanoyl-2-(dimethylamino)naphthalene (Laurdan) confirmed that such effects were restricted to the bilayer surface. Furthermore, the lysolecithin appeared to be the product most responsible for these changes. These results suggested that lysolecithin increases the activity of phospholipase A2 during vesicle hydrolysis by disrupting the bilayer surface, making the phospholipid molecules more accessible to the enzyme active site.
Sheffield
M J
MJ
Department of Zoology, Brigham Young University, Provo, Utah 84602, USA.
Baker
B L
BL
Li
D
D
Owen
N L
NL
Baker
M L
ML
Bell
J D
JD
D015081:2-Naphthylamine / Q000031:analogs & derivatives; D017836:Agkistrodon; D000818:Animals; D003435:Crotalid Venoms / Q000201:enzymology*; D005456:Fluorescent Dyes; D006868:Hydrolysis; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000378:metabolism*; D010741:Phospholipases A / Q000187:drug effects / Q000378:metabolism*; D054467:Phospholipases A2; D013050:Spectrometry, Fluorescence
D003435:Crotalid Venoms; D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D008244:Lysophosphatidylcholines; D010169:Palmitic Acids; D010713:Phosphatidylcholines; D019308:Palmitic Acid; C038517:prodan; D015081:2-Naphthylamine; D010741:Phospholipases A; D054467:Phospholipases A2