title: Purification and characterization of two fibrinolytic enzymes from Bothrops jararaca (jararaca) venom.
abstract: Two fibrinolytic enzymes, jararafibrase I and jararafibrase II, were purified from Bothrops jararaca venom. The purified jararafibrase I and jararafibrase II ran as single protein bands on analytical polyacrylamide gel electrophoresis and had mol. wts of 47,000 +/- 2000 and 21,400 +/- 500, respectively, by SDS-polyacrylamide gel electrophoresis. The isoelectric points of jararafibrase I and jararafibrase II were 4.6 and 6.5, respectively. The specific activities of jararafibrase I and jararafibrase II were 2.2 units/mg protein and 6.3 units/mg protein, respectively. Both enzymes exhibited no detectable plasminogen activating activity. The activity of the enzymes was completely inhibited by 1,10-phenanthroline and ethylenediaminetetraacetate, suggesting that both enzymes were metalloproteinases. Jararafibrase I and jararafibrase II had single-chain protein compositions, and the amino acid sequence up to the 49th amino acid from the NH2-terminal of jararafibrase II was: Leu-Pro-Glu-His-Gln-Arg-Tyr-Ile-Glu-Leu-Phe-Ile-Val-Val-Asp-His-Gly-Met- Phe-Met-Lys-Tyr-Asn-Gly-Asn-Ser-Asp-Lys-Ile-Arg-Arg-Arg-Ile-His-Gln- Met-Val-Asn-Ile-Met-Lys-X-Ala-Tyr-Arg-Tyr-Leu-Tyr-Ile-(X = not confirmed).
authors.affiliation: Department of Physiology, Miyazaki Medical College, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Maruyama
authors.forename: M
authors.initials: M
authors.lastname: Sugiki
authors.forename: E
authors.initials: E
authors.lastname: Yoshida
authors.forename: H
authors.initials: H
authors.lastname: Mihara
authors.forename: N
authors.initials: N
authors.lastname: Nakajima
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003435:Crotalid Venoms / Q000201:enzymology*; D005343:Fibrinolytic Agents / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D012911:Snakes
chemical_list: D003435:Crotalid Venoms; D005343:Fibrinolytic Agents; D008666:Metalloendopeptidases; C076581:jararafibrase I; C076582:jararafibrase II