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 venom of Bothrops jararaca. The protein, named BJ-PLA2, was isolated by means of ammonium sulfate precipitation and anion-exchange and reversed-phase chromatographies and behaved as a homogeneous single-chain protein on SDS-PAGE. Its amino acid sequence was determined by N-terminal sequencing and analysis of overlapped chemical and proteolytic fragments by automated Edman degradation and mass spectometry determination. BJ-PLA2 consists of 124 amino acid residues and has the structural features of snake venom class II phospholipases A2. Chemical modification with p-bromophenacylbromide caused complete loss of enzymatic activity and partially affected the platelet-aggregation-inhibiting activity of BJ-PLA2.
Serrano
S M
SM
Laboratory of Biochemistry and Biophysics and Laboratory of Pathophysiology Instituto Butantan, São Paulo, 05503-900, Brazil. s_serrano@hotmail.com
Reichl
A P
AP
Mentele
R
R
Auerswald
E A
EA
Santoro
M L
ML
Sampaio
C A
CA
Camargo
A C
AC
Assakura
M T
MT
D000098:Acetophenones / Q000378:metabolism; D000244:Adenosine Diphosphate / Q000037:antagonists & inhibitors / Q000494:pharmacology; D000478:Alkylation; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D017837:Bothrops; D011232:Chemical Precipitation; D002853:Chromatography, Liquid; D003094:Collagen / Q000037:antagonists & inhibitors / Q000494:pharmacology; D006801:Humans; D020128:Inhibitory Concentration 50; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010741:Phospholipases A / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D054467:Phospholipases A2; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D017386:Sequence Homology, Amino Acid; D012910:Snake Venoms / Q000201:enzymology*
D000098:Acetophenones; D010975:Platelet Aggregation Inhibitors; D012910:Snake Venoms; D000244:Adenosine Diphosphate; D003094:Collagen; D010741:Phospholipases A; D054467:Phospholipases A2; C006821:4-bromophenacyl bromide