A novel phospholipase A2, BJ-PLA2, from the venom of the snake Bothrops jararaca: Purification, primary structure analysis, and its characterization as a platelet-aggregation-inhibiting factor

This paper describes the isolation and primary structure analysis of a new phospholipase A2 with platelet-aggregation-inhibiting activity from the ven om of Bothrops jararaca. The protein, named BJ-PLA2, was isolated by means of ammonium sulfate precipitation and anion-exchange and reversed-phase chr omatographies and behaved as a homogeneous single-chain protein on SDS-PAGE . Its amino acid sequence was determined by N-terminal sequencing and analy sis of overlapped chemical and proteolytic fragments by automated Edman deg radation and mass spectometry determination. BJ-PLA2 consists of 124 amino acid residues and has the structural features of snake venom class II phosp holipases A2, Chemical modification with p-bromophenacylbromide caused comp lete loss of enzymatic activity and partially affected the platelet-aggrega tion-inhibiting activity of BJ-PLA2. (C) 1999 Academic Press.