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Isolation and properties of two phospholipases A2 from the venom of an Australian elapid snake (Pseudechis australis).

Abstract
Two phospholipases A2 (Pa-11 and Pa-13) were purified from the venom of an Australian elapid snake (subfamily Acanthophiinae) Pseudechis australis (king brown snake) by chromatography on CM-cellulose CM-52 followed by gel filtration on a Sephadex G-75 column. The apparent molecular weights of the two phospholipases A2 (Pa-11 and Pa-13) were 14,000 and 13,500, respectively, by gel filtration analysis on a Sephadex G-75 column. Each enzyme molecule consists of a single polypeptide chain of 118 amino acid residues. The isoelectric points of Pa-11 and Pa-13 were 10.5 and 10.0, respectively. The optimum pH values of Pa-11 and Pa-13 for hydrolysis of egg-yolk phosphatidylcholine were 7.8 and 7.5, respectively. Pa-11 was lethal to mice (LD50 0.23 micrograms/g body weight), whereas Pa-13 showed no lethal activity at a dose level of 7.4 micrograms/g mouse. Each enzyme was inactivated by reaction with p-bromophenacylbromide on the sole histidine residue (Pa-11) or on one of the two histidine residues (Pa-13). Oxidation of the tryptophan residues in Pa-11 and Pa-13 with N-bromosuccinimide led to a decrease in the phospholipase A activity. A complete loss of both enzymic and lethal activities of Pa-11 was observed upon oxidation of one of the two tryptophan residues of the molecule.
AuthorsS Nishida, M Terashima, T Shimazu, C Takasaki, N Tamiya
JournalToxicon : official journal of the International Society on Toxinology (Toxicon) Vol. 23 Issue 1 Pg. 73-85 ( 1985) ISSN: 0041-0101 [Print] England
PMID3992600 (Publication Type: Journal Article)
Chemical References
  • Acetophenones
  • Amino Acids
  • Elapid Venoms
  • Pseudechis venom
  • Phospholipases
  • Phospholipases A
  • Bromosuccinimide
  • 4-bromophenacyl bromide
Topics
  • Acetophenones
  • Amino Acids (analysis)
  • Animals
  • Bromosuccinimide
  • Chromatography, Gel
  • Crystallization
  • Elapid Venoms (analysis)
  • Electrophoresis, Polyacrylamide Gel
  • Isoelectric Focusing
  • Lethal Dose 50
  • Mice
  • Molecular Weight
  • Oxidation-Reduction
  • Phospholipases (isolation & purification)
  • Phospholipases A (isolation & purification, metabolism, toxicity)

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