Characterization of phospholipase A2 from rabbit lung microsomes.
A phospholipase A2 activity associated with the microsomal fraction of rabbit lung homogenates was studied. The enzyme showed specificity for the sn-2 ester bond of phosphatidylcholine, had an alkaline pH optimum and required Ca2+ for activity. Other divalent cations were unable to support hydrolysis. In the absence of detergents, exogenous phosphatidylethanolamine was deacylated at a rate sevenfold higher than phosphatidylcholine. The activity toward both substrates could be enhanced by sodium deoxycholate or, more effectively, by sodium taurodeoxycholate. Phosphatidylethanolamine required higher detergent/phospholipid molar ratios than phosphatidylcholine. Under these conditions, the preference for the former substrate over the latter was nearly abolished. The zwitterionic detergent 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS) and the nonionic detergent Triton X-100 were either ineffective (phosphatidylcholine) or inhibitory (phosphatidylethanolamine). Addition of KCl produced opposite effects on the activity depending on the bile salt used to disperse the substrate. The phospholipase A2 activity was inhibited by p-bromophenacyl bromide but remained unaffected after treatment with diisopropylfluorophosphate or NaF. N-Ethylmaleimide, but not other thiol reagents, partially inhibited the activity.
Department of Obstetrics and Gynecology, University of Western Ontario, London, Canada.
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D000818:Animals; D003902:Detergents / Q000494:pharmacology; D005260:Female; D008168:Lung / Q000201:enzymology*; D008563:Membrane Lipids / Q000032:analysis; D008861:Microsomes / Q000201:enzymology*; D010713:Phosphatidylcholines / Q000378:metabolism; D010740:Phospholipases / Q000302:isolation & purification*; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D054467:Phospholipases A2; D010743:Phospholipids / Q000096:biosynthesis; D011189:Potassium Chloride / Q000494:pharmacology; D011817:Rabbits; D013438:Sulfhydryl Compounds / Q000494:pharmacology
D003902:Detergents; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010743:Phospholipids; D013438:Sulfhydryl Compounds; D011189:Potassium Chloride; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2