title: Isolation, properties and amino acid sequences of a phospholipase A2 and its homologue without activity from the venom of a sea snake, Laticauda colubrina, from the Solomon Islands.
abstract: A phospholipase A2, Laticauda colubrina phospholipase A2 II (LcPLA-II), and a phospholipase A2 homologue, Laticauda colubrina phospholipase A2 homologue I (LcPLH-I), were isolated from the venom of the yellow-lipped sea snake, Laticauda colubrina, from the Solomon Islands. LcPLA-II showed phospholipase A2 activity towards egg-yolk phosphatidylcholine (24 mumol/min per mg at optimal conditions at 37 degrees C) and lethal potency (LD50 45 micrograms/kg body wt. intravenously in mice). Both of the activities were lost by treatment with p-bromophenacyl bromide. LcPLH-I showed neither phospholipase A2 activity nor lethal potency at a dose of 4.5 mg/kg body wt. in mice. It was not modified by the treatment with p-bromophenacyl bromide. LcPLA-II and LcPLH-I bound Ca2+ at a 1:1 molar ratio with KCa values of 105 microM and 44 microM at pH 8.0 respectively. Elucidation of the amino acid sequences of these two proteins showed that each protein consisted of a single chain of 118 amino acid residues, including 14 half-cystine residues. The two sequences are different from each other at 22 residues and highly homologous to those from other sources. The essential histidine residue for the phospholipase A2 activity at position 48 is replaced by an asparagine residue in the homologue LcPLH-I. Details of the separation of the peptides obtained by proteinase digestions of LcPLA-II and LcPLA-I and the determination of their amino acid sequences are given in Supplementary Publication SUP 50145 (14 pages), which has been deposited at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1988) 249, 5.
authors.affiliation: Department of Chemistry, Faculty of Science, Tohoku University, Sendai, Japan.
authors.forename: C
authors.initials: C
authors.lastname: Takasaki
authors.forename: S
authors.initials: S
authors.lastname: Kimura
authors.forename: Y
authors.initials: Y
authors.lastname: Kokubun
authors.forename: N
authors.initials: N
authors.lastname: Tamiya
mesh_terms: D000098:Acetophenones / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D004546:Elapid Venoms / Q000032:analysis*; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007928:Lethal Dose 50; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D010740:Phospholipases / Q000378:metabolism*; D010741:Phospholipases A / Q000302:isolation & purification / Q000378:metabolism* / Q000633:toxicity; D054467:Phospholipases A2; D012689:Sequence Homology, Nucleic Acid
chemical_list: D000098:Acetophenones; D004546:Elapid Venoms; D007527:Isoenzymes; D010446:Peptide Fragments; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; C006821:4-bromophenacyl bromide