title: Separation of monomerizing and lysozyme activities of destabilase from medicinal leech salivary gland secretion.
abstract: Destabilase, endo-epsilon-(gamma-Glu)-Lys-isopeptidase, was prepared from the salivary gland secretion of the medicinal leech (Hirudo medicinalis). The secretion prepared by the known method of Rigbi et al. (1987) (secretion-K) lacks the destabilase-characteristic highly specific isopeptidase activity (the D-dimer-monomerizing activity) because of its degradation by proteolytic activity (the substrate of Glp-Ala-Ala-Leu-pNA) due to contamination with leech intestinal channel contents. Therefore, we have elaborated a new technique for preparation of a true leech secretion (secretion-I). This secretion is characterized by the complete absence of the leech intestinal channel contents and has no proteolytic activity. For the first time the destabilase-specific D-dimer-monomerizing and lysozyme activities were separated by fractionation of secretion-I by HPLC gel filtration through Superose S-12. For the purified destabilase preparation, these activities were separated by reversed-phase chromatography in an acetonitrile gradient (0-60%) in the presence of 0.1% trifluoroacetic acid. The monomerizing activity of destabilase is responsible for the ability of secretion-I to dissolve stabilized fibrin via isopeptidolysis of alpha-alpha and gamma-gamma fibrin chains bound by epsilon-(gamma-Glu)-Lys-isopeptide bonds.
authors.affiliation: School of Biology, Lomonosov Moscow State University, Moscow, 119899 Russia. leech@humgen.siobc.ras.ru
authors.forename: I P
authors.initials: IP
authors.lastname: Baskova
authors.forename: L L
authors.initials: LL
authors.lastname: Zavalova
authors.forename: A V
authors.initials: AV
authors.lastname: Basanova
authors.forename: A V
authors.initials: AV
authors.lastname: Sass
mesh_terms: D000818:Animals; D010450:Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism; D005337:Fibrin / Q000378:metabolism*; D007865:Leeches / Q000201:enzymology*; D009113:Muramidase / Q000378:metabolism*; D012469:Salivary Glands / Q000201:enzymology* / Q000378:metabolism
chemical_list: D005337:Fibrin; D009113:Muramidase; D010450:Endopeptidases; C045348:fibrin destabilase