Identification of proteases in the extract of venom glands from brown spiders

Identification of proteases in the extract of venom glands from brown spiders

Author Silveira, R. B. da Google Scholar
Santos, J. F. dos Google Scholar
Mangili, O. C. Google Scholar
Veiga, S. S. Google Scholar
Gremski, W. Google Scholar
Nader, H. B. Google Scholar
von Dietrich, C. P. Google Scholar
Institution Univ Fed Parana
Universidade Federal de São Paulo (UNIFESP)
Abstract In the present investigation, in order to dispute the rational criticism against the presence of proteolytic enzymes in the electrostimulated venom obtained from spiders of the genus Loxosceles, as a consequence of contamination with abdominal secretions, venoms of L. intermedia and L. laeta were directly collected from venom glands by microdissection and gentle homogenization. Gel electrophoresis stained by silver method carried out to compare L. intermedia electrostimulated venom and venom gland extract demonstrated no significant differences in protein profile. Zymogram analysis of L. intermedia venom gland extract detected a gelatinolytic activity in the 32-35 kDa region. the inhibitory effect of 1,10-phenanthroline on this proteolytic activity further supported its metalloprotease nature. in proteolytic digestion experiments L. intermedia venom gland extract was also able to cleave purified fibronectin and fibrinogen. the inhibitory effect of 1, 10-phenanthroline on these degrading activities confirmed the presence of metalloproteases in the venom. in addition, when purified fibrinogen was incubated with L. intermedia abdominal extract, the fibrinogenolysis was completely different, generating low mass fragments that ran away from the gel, a proteolytic event not blocked by 1,10-phenanthroline. Zymogram experiments using L. laeta venom gland extracts further detected a gelatinolytic band at 32-35 kDa, also inhibited by 1,10-phenanthroline, confirming the presence of metalloproteases in both species. (C) 2002 Elsevier B.V. All rights reserved.
Keywords proteases
Loxosceles
zymogram
Language English
Date 2002-06-01
Published in Toxicon. Oxford: Pergamon-Elsevier B.V., v. 40, n. 6, p. 815-822, 2002.
ISSN 0041-0101 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 815-822
Origin http://dx.doi.org/10.1016/S0041-0101(02)00078-8
Access rights Closed access
Type Article
Web of Science ID WOS:000175810700019
URI http://repositorio.unifesp.br/handle/11600/26883

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