title: Rapid purification of serine proteinases from Bothrops alternatus and Bothrops moojeni venoms.
abstract: Envenomation by Bothrops species results, among other symptoms, in hemostatic disturbances. These changes can be ascribed to the presence of enzymes, primarily serine proteinases some of which are structurally similar to thrombin and specifically cleave fibrinogen releasing fibrinopeptides. A rapid, three-step, chromatographic procedure was developed to routinely purify serine proteinases from the venoms of Bothrops alternatus and Bothrops moojeni. The serine proteinase from B. alternatus displays an apparent molecular mass of ~32 kDa whereas the two closely related serine proteinases from B. moojeni display apparent molecular masses of ~32 kDa and ~35 kDa in SDS-PAGE gels. The partial sequences indicated that these enzymes share high identity with serine proteinases from the venoms of other Bothrops species. These proteins coagulate plasma and possess fibrinogenolytic activity but lack fibrinolytic activity.
authors.affiliation: Centro Multiusuário de Inovação Biomolecular, Departamento de Física, Universidade Estadual Paulista (UNESP), São José do Rio Preto, SP 15054-000, Brazil.
authors.forename: Liliane Maria
authors.initials: LM
authors.lastname: Fernandes de Oliveira
authors.forename: Anwar
authors.initials: A
authors.lastname: Ullah
authors.forename: Rehana
authors.initials: R
authors.lastname: Masood
authors.forename: André
authors.initials: A
authors.lastname: Zelanis
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Spencer
authors.forename: Solange M T
authors.initials: SM
authors.lastname: Serrano
authors.forename: Raghuvir K
authors.initials: RK
authors.lastname: Arni
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D017837:Bothrops; D003435:Crotalid Venoms / Q000201:enzymology*; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010949:Plasma / Q000187:drug effects; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D057057:Serine Proteases / Q000302:isolation & purification* / Q000494:pharmacology
chemical_list: D003435:Crotalid Venoms; D057057:Serine Proteases
keywords: Bothrops alternatus; Bothrops moojeni; Crude venom; Fibrinogenolysis; MCD; PAGE; Proteolytic activity; SDS; SVSPs; Serine proteinases; minimum coagulant dose; polyacrylamide gel electrophoresis; snake venom serine proteinases; sodium dodecyl sulfate