Isolation and properties of lysophospholipases from the venom of an Australian elapid snake, Pseudechis australis.
Two lysophospholipases were isolated from the venom of an Australian elapid snake (subfamily Acanthophiinae), Pseudechis australis, by sequential chromatography on CM-52 cellulose, Sephadex G-75 and DE-52 cellulose columns. They were very similar to each other. One of them, lysophospholipase I, was obtained as a homodimer, the monomer of which consisted of 123 amino acid residues with seven disulphide bridges. The amino acid composition and the N-terminal amino acid sequence of the enzyme were similar to those of phospholipase A2, Ca2+ was required for its activity and the maximum activity was attained at 2 mM-CaCl2 in the presence of 1 mM-EDTA. The optimum pH was 7.5. Lysophospholipase I hydrolysed lysophosphatidylcholine more rapidly than lysophosphatidylethanolamine. It did not hydrolyse, however, phosphatidylcholine, 1-palmitoylglycerol, tripalmitoylglycerol or p-nitrophenyl acetate. Modification of the enzyme with p-bromophenacyl bromide or 2-nitrophenylsulphenyl chloride suppressed the activity. A strong direct haemolytic activity was exhibited when the lysophospholipase was present together with phospholipase A2.
D000098:Acetophenones / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D004546:Elapid Venoms; D006868:Hydrolysis; D007202:Indicators and Reagents / Q000494:pharmacology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D008244:Lysophosphatidylcholines / Q000378:metabolism; D008245:Lysophospholipase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D009578:Nitrobenzenes / Q000494:pharmacology; D010740:Phospholipases / Q000378:metabolism*; D013379:Substrate Specificity
D000098:Acetophenones; D004546:Elapid Venoms; D007202:Indicators and Reagents; D007527:Isoenzymes; D008244:Lysophosphatidylcholines; D009578:Nitrobenzenes; C002732:2-nitrobenzenesulfenyl chloride; D010740:Phospholipases; D008245:Lysophospholipase; C006821:4-bromophenacyl bromide; D002118:Calcium