Purification and primary structure of a myotoxic Lysine-49 phospholipase A2 with low lipolytic activity from Trimeresurus gramineus venom

Makoto Nakai, Kin Ichi Nakashima, Tomohisa Ogawa, Yasuyuki Shimohigashi, Shosaku Hattori, Chun Chang Chang, Motonori Ohno

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Abstract

Four acidic phospholipase A2 (PLA2) isozymes named PLA2-I, II, III and IV have previously been isolated from Trimeresurus gramineus (green habu snake) venom and sequenced [Oda et al. (1991) Toxicon29, 157; Fukagawa et al. (1992) Toxicon30, 1131; Fukagawa et al. (1993) Toxicon 31, 957]. They contain aspartate-49 which is known to bind Ca2+, essential for catalysis. In the present study, a basic PLA2 named PLA2-V containing lysine-49 was newly isolated from the same snake venom. Its isoelectric point was 9.4 and considerably higher than those (c. 4.5) of PLA2-I-IV. PLA2-V was 1.1% as active as PLA2-I toward egg-yolk emulsion but exhibited strong myotoxicity. The amino acid sequence of PLA2-V was determined by sequencing the S-carboxamidomethylated derivative and its peptide fragments produced by enzymatic (clostripain, chymotrypsin, Achromobacter protease I and Staphylococcus aureus V8 protease) cleavages. PLA2-V consists of 122 amino acid residues and is highly homologous (72-78%) to Lys-49 PLA2s so far isolated from Viperidae snake venoms but less homologous (52%) to PLA2-I. The presence of Asn-28, which is characteristic of Lys-49 PLA2s, was confirmed.

Original languageEnglish
Pages (from-to)1469-1478
Number of pages10
JournalToxicon
Volume33
Issue number11
DOIs
Publication statusPublished - Jan 1 1995
Externally publishedYes

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All Science Journal Classification (ASJC) codes

  • Toxicology

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