title: Isolation, functional, and partial biochemical characterization of galatrox, an acidic lectin from Bothrops atrox snake venom.
abstract: Snake venom lectins have been studied in regard to their chemical structure and biological functions. However, little is known about lectins isolated from Bothrops atrox snake venom. We report here the isolation and partial functional and biochemical characterization of an acidic glycan-binding protein called galatrox from this venom. This lectin was purified by affinity chromatography using a lactosyl-sepharose column, and its homogeneity and molecular mass were evaluated by high-performance liquid chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The purified galatrox was homogeneous and characterized as an acidic protein (pI 5.2) with a monomeric and dimeric molecular mass of 16.2 and 32.5 kDa, respectively. Alignment of N-terminal and internal amino acid sequences of galatrox indicated that this protein exhibits high homology to other C-type snake venom lectins. Galatrox showed optimal hemagglutinating activity at a concentration of 100 μg/ml and this effect was drastically inhibited by lactose, ethylenediaminetetraacetic acid, and heating, which confirmed galatrox's lectin activity. While galatrox failed to induce the same level of paw edema or mast cell degranulation as B. atrox crude venom, galatrox did alter cellular viability, which suggested that galatrox might contribute to venom toxicity by directly inducing cell death.
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil.
authors.forename: Elaine de Paula
authors.initials: Ede P
authors.lastname: Mendonça-Franqueiro
authors.forename: Raquel de Melo
authors.initials: Rde M
authors.lastname: Alves-Paiva
authors.forename: Marco Aurélio
authors.initials: MA
authors.lastname: Sartim
authors.forename: Daniel Roberto
authors.initials: DR
authors.lastname: Callejon
authors.forename: Helder Henrique
authors.initials: HH
authors.lastname: Paiva
authors.forename: Gilmara Ausech
authors.initials: GA
authors.lastname: Antonucci
authors.forename: José César
authors.initials: JC
authors.lastname: Rosa
authors.forename: Adélia Cristina Oliveira
authors.initials: AC
authors.lastname: Cintra
authors.forename: João José
authors.initials: JJ
authors.lastname: Franco
authors.forename: Eliane Candiani
authors.initials: EC
authors.lastname: Arantes
authors.forename: Marcelo
authors.initials: M
authors.lastname: Dias-Baruffi
authors.forename: Suely Vilela
authors.initials: SV
authors.lastname: Sampaio
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops / Q000378:metabolism*; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line / Q000166:cytology / Q000187:drug effects; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004487:Edema / Q000139:chemically induced; D037102:Lectins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism / Q000633:toxicity; D008297:Male; D008407:Mast Cells / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D051381:Rats; D017386:Sequence Homology, Amino Acid; D012910:Snake Venoms / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism / Q000633:toxicity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D014757:Viper Venoms / Q000737:chemistry* / Q000302:isolation & purification* / Q000633:toxicity
chemical_list: D037102:Lectins; D012910:Snake Venoms; D014757:Viper Venoms; C557501:galatrox protein, Bothrops atrox