title: A New Phospholipase A₂ from Lachesis muta rhombeata: Purification, Biochemical and Comparative Characterization with Crotoxin B.
abstract: Phospholipases A2 (PLA2s) are enzymes responsible for inflammatory effects, edema formation, myotoxicity, neurotoxicity and other manifestations from envenoming. In this paper we report the isolation and biochemical characterization of Lmr-PLA2, the first acidic PLA2 found in Lachesis muta rhombeata venom. Furthermore, this study compared biological effects of Lmr-PLA2 and crotoxin B (CB), a PLA2 from Crotalus durissus terrificus venom. Lmr-PLA2 was isolated by molecular exclusion and reversed phase chromatography. The purified enzyme showed a molecular mass of 13,975 Da, pI of 5.46 and its partial amino acid sequence showed a high identity with PLA2s already described in the literature. In addition, this enzyme possesses the residue D49 in its amino acid sequence, indicating that it is a catalytically active PLA2. Lmr-PLA2 presented high phospholipase activity and was able to inhibit platelet aggregation. Studies of biochemical characterization of new PLA2s, as Lmr-PLA2, are relevant since they help to clarify the structure-function relationship of this important class of toxins.
authors.forename: Francielle A
authors.initials: FA
authors.lastname: Cordeiro
authors.forename: Tibério G K
authors.initials: TG
authors.lastname: Perini
authors.forename: Cristiane
authors.initials: C
authors.lastname: Bregge-Silva
authors.forename: Caroline M
authors.initials: CM
authors.lastname: Cremonez
authors.forename: Renata S
authors.initials: RS
authors.lastname: Rodrigues
authors.forename: Johara
authors.initials: J
authors.lastname: Boldrini-França
authors.forename: Karla de C F
authors.initials: Kde C
authors.lastname: Bordon
authors.forename: Dayane L N
authors.initials: DL
authors.lastname: De Souza
authors.forename: David C
authors.initials: DC
authors.lastname: Ache
authors.forename: Veridiana
authors.initials: V
authors.lastname: de M Rodrigues
authors.forename: Wagner F
authors.initials: WF
authors.lastname: Dos Santos
authors.forename: Jose C
authors.initials: JC
authors.lastname: Rosa
authors.affiliation: Universidade de São Paulo, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Departamento de Física e Química Av. do Café, s/n, 14040-903, Ribeirão Preto-SP, Brazil. ecabraga@fcfrp.usp.br.
authors.forename: Eliane C
authors.initials: EC
authors.lastname: Arantesa
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003402:Creatine Kinase / Q000032:analysis / Q000378:metabolism; D003439:Crotoxin / Q000737:chemistry; D004487:Edema / Q000139:chemically induced; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000187:drug effects / Q000473:pathology; D010446:Peptide Fragments / Q000032:analysis; D054467:Phospholipases A2 / Q000737:chemistry* / Q000302:isolation & purification* / Q000633:toxicity; D010974:Platelet Aggregation / Q000187:drug effects; D030162:Reptilian Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000633:toxicity; D016415:Sequence Alignment; D014757:Viper Venoms / Q000737:chemistry*; D017819:Viperidae
chemical_list: D010446:Peptide Fragments; D030162:Reptilian Proteins; D014757:Viper Venoms; D003439:Crotoxin; D003402:Creatine Kinase; D054467:Phospholipases A2