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Further characterization and N-terminal sequence of cobra venom phospholipase A2

1980; Elsevier BV; Volume: 626; Issue: 1 Linguagem: Inglês

10.1016/0005-2795(80)90206-8

1878-1454

Paul L. Darke, Alice A. Jarvis, Raymond A. Deems, Edward A. Dennis,

Insect and Pesticide Research

N-terminal amino acid sequence data on cobra venom phospholipase A2 (Naja naja naja) demonstrate the absence of histidine at positions 10 and 20, in contrast to most other Naja enzymes. The presence of three tryptophans was demonstrated by p-toluenesulfonic acid hydrolysis, and the location of two of these at positions 18 and 19 parallels other Naja phospholipase A2 sequences. Based on the amino acid composition of the enzyme, the theoretical E2780.1% should be 2.2, which is consistent with dry weight determinations, refractometry and amino acid analysis. Unusually high reactivity of the enzyme toward the protein assay of Lowry et al. gives an erroneously low E2780.1% value of 1.45 when compared to bovine serum serum albumin as a protein standard. Using an E2780.1% of 2.2, this enzyme is inactivated by the active site reagent p-bromophenacyl bromide with a stoichiometry of 1 mol reagent per mol enzyme. Chromatography on Affi-Gel Blue provides further purification of this phospholipase A2.