title: Snake venomics and antivenomics of the arboreal neotropical pitvipers Bothriechis lateralis and Bothriechis schlegelii.
abstract: We report the comparative proteomic characterization of the venoms of two related neotropical arboreal pitvipers from Costa Rica of the genus Bothriechis, B. lateralis (side-striped palm pit viper) and B. schlegelii (eyelash pit viper). The crude venoms were fractionated by reverse-phase HPLC, followed by analysis of each chromatographic fraction by SDS-PAGE, N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry of tryptic peptides. The venom proteomes of B. lateralis and B. schlegelii comprise similar number of distinct proteins belonging, respectively, to 8 and 7 protein families. The two Bothriechis venoms contain bradykinin-potentiating peptides (BPPs), and proteins from the phospholipase A 2 (PLA 2), serine proteinase, l-amino acid oxidase (LAO), cysteine-rich secretory protein (CRISP), and Zn (2+)-dependent metalloproteinase (SVMP) families, albeit each species exhibit different relative abundances. Each venom also contains unique components, for example, snake venom vascular endothelial growth factor (svVEGF) and C-type lectin-like molecules in B. lateralis, and Kazal-type serine proteinase inhibitor-like proteins in B. schlegelii. Using a similarity coefficient, we estimate that the similarity of the venom proteins between the two Bothriechis taxa may be <10%, indicating a high divergence in their venom compositions, in spite of the fact that both species have evolved to adapt to arboreal habits. The major toxin families of B. lateralis and B. schlegelii are SVMP (55% of the total venom proteins) and PLA 2 (44%), respectively. Their different venom toxin compositions provide clues for rationalizing the distinct signs of envenomation caused by B. schlegelii and B. lateralis. An antivenomic study of the immunoreactivity of the Instituto Clodomiro Picado (ICP) polyvalent antivenom toward Bothriechis venoms revealed that l-amino acid oxidase and SVMPs represent the major antigenic protein species in both venoms. Our results provide a ground for rationalizing the reported protection of the ICP polyvalent antivenom against the hemorrhagic, coagulant, defibrinating, caseinolytic and fibrin(ogen)olytic activities of Bothriechis ( schlegelii, lateralis) venoms. However, these analyses also evidenced the limited recognition capability of the polyvalent antivenom toward a number of Bothriechis venom components, predominantly BPPs, svVEGF, Kazal-type inhibitors, some PLA 2 proteins, some serine proteinases, and CRISP molecules.
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
authors.forename: Bruno
authors.initials: B
authors.lastname: Lomonte
authors.forename: José
authors.initials: J
authors.lastname: Escolano
authors.forename: Julián
authors.initials: J
authors.lastname: Fernández
authors.forename: Libia
authors.initials: L
authors.lastname: Sanz
authors.forename: Yamileth
authors.initials: Y
authors.lastname: Angulo
authors.forename: José María
authors.initials: JM
authors.lastname: Gutiérrez
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Calvete
mesh_terms: D000818:Animals; D000997:Antivenins / Q000032:analysis* / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D003435:Crotalid Venoms / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D050830:L-Amino Acid Oxidase / Q000032:analysis / Q000276:immunology; D037181:Lectins, C-Type / Q000032:analysis; D045726:Metalloproteases / Q000032:analysis / Q000276:immunology; D009842:Oligopeptides / Q000032:analysis; D054497:Phospholipases A2, Secretory / Q000032:analysis; D020543:Proteome / Q000032:analysis* / Q000276:immunology; D012697:Serine Endopeptidases / Q000032:analysis; D015842:Serine Proteinase Inhibitors / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis; D017819:Viperidae / Q000378:metabolism*
chemical_list: D000997:Antivenins; D003435:Crotalid Venoms; D037181:Lectins, C-Type; D009842:Oligopeptides; D020543:Proteome; D015842:Serine Proteinase Inhibitors; D042461:Vascular Endothelial Growth Factor A; C006791:bradykinin potentiating factors; D050830:L-Amino Acid Oxidase; D054497:Phospholipases A2, Secretory; D045726:Metalloproteases; D012697:Serine Endopeptidases