title: Isolation and functional characterization of proinflammatory acidic phospholipase A2 from Bothrops leucurus snake venom.
abstract: In the present study, an acidic PLA(2), designated Bl-PLA(2), was isolated from Bothrops leucurus snake venom through two chromatographic steps: ion-exchange on CM-Sepharose and hydrophobic chromatography on Phenyl-Sepharose. Bl-PLA(2) was homogeneous on SDS-PAGE and when submitted to 2D electrophoresis the molecular mass was 15,000Da and pI was 5.4. Its N-terminal sequence revealed a high homology with other Asp49 acidic PLA(2)s from snake venoms. Its specific activity was 159.9U/mg and the indirect hemolytic activity was also higher than that of the crude venom. Bl-PLA(2) induced low myotoxic and edema activities as compared to those of the crude venom. Moreover, the enzyme was able to induce increments in IL-12p40, TNF-α, IL-1β and IL-6 levels and no variation of IL-8 and IL-10 in human PBMC stimulated in vitro, suggesting that Bl-PLA(2) induces proinflammatory cytokine production by human mononuclear cells. Bothrops leucurus venom is still not extensively explored and knowledge of its components will contribute for a better understanding of its action mechanism.
authors.affiliation: Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, UFU, Uberlândia, MG, Brazil.
authors.forename: Débora C O
authors.initials: DC
authors.lastname: Nunes
authors.forename: Renata S
authors.initials: RS
authors.lastname: Rodrigues
authors.forename: Malson N
authors.initials: MN
authors.lastname: Lucena
authors.forename: Camila T
authors.initials: CT
authors.lastname: Cologna
authors.forename: Ana Carolina S
authors.initials: AC
authors.lastname: Oliveira
authors.forename: Amélia
authors.initials: A
authors.lastname: Hamaguchi
authors.forename: Maria I
authors.initials: MI
authors.lastname: Homsi-Brandeburgo
authors.forename: Eliane C
authors.initials: EC
authors.lastname: Arantes
authors.forename: David N S
authors.initials: DN
authors.lastname: Teixeira
authors.forename: Carlos
authors.initials: C
authors.lastname: Ueira-Vieira
authors.forename: Veridiana M
authors.initials: VM
authors.lastname: Rodrigues
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops; D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange / Q000379:methods; D003435:Crotalid Venoms / Q000201:enzymology*; D016207:Cytokines / Q000378:metabolism; D004487:Edema / Q000139:chemically induced; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007249:Inflammation / Q000378:metabolism*; D016753:Interleukin-10 / Q000378:metabolism; D053762:Interleukin-12 Subunit p40 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008970:Molecular Weight; D054467:Phospholipases A2 / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D012685:Sepharose / Q000031:analogs & derivatives; D017386:Sequence Homology, Amino Acid; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D003435:Crotalid Venoms; D016207:Cytokines; D053762:Interleukin-12 Subunit p40; D015850:Interleukin-6; D016209:Interleukin-8; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; C020927:phenyl-sepharose; D012685:Sepharose; D054467:Phospholipases A2