title: Purification, physicochemical characterization and N-terminal-amino acid sequence of a phospholipase A2 from Bothrops jararaca venom.
abstract: Snake venoms usually contain multiple molecular forms of phospholipase A2 enzymes (phosphatide acyl hydrolase, E.C. 3.1.1.4; PLA2). Phospholipases A2 induce a wide range of pharmacological effects which may depend or not on the hydrolysis of phospholipids. In this study, a PLA2 from Bothrops jararaca venom was purified to homogeneity by gel filtration on a Sephacryl S-200 column, followed by FPLC reverse-phase chromatography on a Pep-RPC HR 5/5 column (yield 1.63% of venom protein). The PLA2 activity of the fractions was determined by indirect hemolysis using hen's egg yolk lecithin as substrate. The enzyme is an acidic protein with PI 4.5 and an apparent molecular weight of 14,200, as estimated by gel filtration on a Superose 12 FPLC column. Similar properties have been described for PLA2 from other snake venoms. The N-terminal-sequence of the purified protein was NLMQFETMIMXXAGQ. These partial sequence data show a high degree of homology between the B. jararaca PLA2 and the enzymes from other snake venoms as well as bovine pancreatic PLA2.
authors.affiliation: Departamento de Bioquímica Médica, Centro de Ciências da Saúde, Rio de Janeiro, Brasil.
authors.forename: O L
authors.initials: OL
authors.lastname: Machado
authors.forename: A L
authors.initials: AL
authors.lastname: Oliveira-Carvalho
authors.forename: R B
authors.initials: RB
authors.lastname: Zingali
authors.forename: C R
authors.initials: CR
authors.lastname: Carlini
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops; D002845:Chromatography; D002850:Chromatography, Gel; D003435:Crotalid Venoms / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010741:Phospholipases A / Q000302:isolation & purification*; D054467:Phospholipases A2; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D003435:Crotalid Venoms; D010741:Phospholipases A; D054467:Phospholipases A2