title: Two related thrombin-like enzymes present in Bothrops atrox venom.
abstract: This article describes the presence of two new forms of a thrombin-like enzyme, both with apparent molecular masses of 38 kDa, in Bothrops atrox venom. Both share the ability to cleave fibrinogen into fibrin and to digest casein. Both present identical K(m) on the substrate BApNA. Their N-terminal amino acid sequences are identical for 26 residues, sharing 80% homology with batroxobin and flavoxobin. Two groups of monoclonal antibodies (mAbs) raised against the purified enzyme forms recognized different epitopes of the putative corresponding enzymes present in B. atrox crude venom. On Western blotting analysis of B. atrox crude venom, mAbs 5DB2C8, 5AA10 and 5CF11, but not mAbs 6CC5 and 6AD2-G5, revealed two or more protein bands ranging from 25 to 38 kDa. By immunoprecipitation assays, the 6AD2-G5 mAb was able to precipitate protein bands of 36-38 kDa from B. atrox, B. leucurus, B. pradoi, B. moojeni, B. jararaca and B. neuwiedii crude venoms. Fibrinogen-clotting activity was inhibited when the same venom specimens were pre-incubated with mAb 6AD2-G5, except for B. jararaca and B. neuwiedii.
authors.affiliation: Laboratório de Biologia do Reconhecer, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ, Brasil.
authors.forename: J H
authors.initials: JH
authors.lastname: Petretski
authors.forename: M
authors.initials: M
authors.lastname: Kanashiro
authors.forename: C P
authors.initials: CP
authors.lastname: Silva
authors.forename: E W
authors.initials: EW
authors.lastname: Alves
authors.forename: T L
authors.initials: TL
authors.lastname: Kipnis
mesh_terms: D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects*; D015153:Blotting, Western; D017837:Bothrops; D003435:Crotalid Venoms / Q000201:enzymology* / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000737:chemistry*; D007527:Isoenzymes / Q000302:isolation & purification / Q000494:pharmacology; D011233:Precipitin Tests; D013917:Thrombin / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D003435:Crotalid Venoms; D007527:Isoenzymes; D005340:Fibrinogen; D013917:Thrombin