Description of a serpin toxin in Loxosceles (Brown spider) venoms: Cloning, expression in baculovirus-infected insect cells and functional characterization.

Autor: Schemczssen-Graeff Z; Department of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, PR, Brazil., Justa HCD; Department of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, PR, Brazil., Nowatzki J; Department of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, PR, Brazil., Baldissera AB; Department of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, PR, Brazil., Polli NLC; Department of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, PR, Brazil., De-Bona E; Department of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, PR, Brazil., Rossi IV; Department of Biochemistry, Federal University of Paraná, (UFPR), Curitiba 81530-900, PR, Brazil., Ramirez MI; Department of Biochemistry, Federal University of Paraná, (UFPR), Curitiba 81530-900, PR, Brazil; Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro 21040-900, RJ, Brazil., Minozzo JC; Center for Production and Research of Immunobiological Products (CPPI), State Department of Health, Piraquara 83302-200, PR, Brazil., Matsubara FH; Department of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, PR, Brazil., Senff-Ribeiro A; Department of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, PR, Brazil., Gremski LH; Department of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, PR, Brazil., Veiga SS; Department of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, PR, Brazil. Electronic address: veigass@ufpr.br.
Jazyk: angličtina
Zdroj: International journal of biological macromolecules [Int J Biol Macromol] 2021 Jul 31; Vol. 183, pp. 1607-1620. Date of Electronic Publication: 2021 May 21.
DOI: 10.1016/j.ijbiomac.2021.05.129
Abstrakt: Several classes of toxins are present in the venom of Brown spiders (Loxosceles genus), some of them are highly expressed and others are less expressed. In this work, we aimed to clone the sequence of a little expressed novel toxin from Loxosceles venom identified as a serine protease inhibitor (serpin), as well as to express and characterize its biochemical and biological properties. It was named LSPILT, derived from Loxoscelesserine protease inhibitor-like toxin. Multiple alignment analysis revealed high identity between LSPILT and other serpin molecules from spiders and crab. LSPILT was produced in baculovirus-infected insect cells, resulting in a 46-kDa protein fused to a His-tag. Immunological assays showed epitopes in LSPILT that resemble native venom toxins of Loxosceles spiders. The inhibitory activity of LSPILT on trypsin was found both by reverse zymography and fluorescent gelatin-degradation assay. Additionally, LSPILT inhibited the complement-dependent lysis of Trypanosoma cruzi epimastigotes, reduced thrombin-dependent clotting and suppressed B16-F10 melanoma cells migration. Results described herein prove the existence of conserved serpin-like toxins in Loxosceles venoms. The availability of a recombinant serpin enabled the determination of its biological and biochemical properties and indicates potential applications in future studies regarding the pathophysiology of the envenoming or for biotechnological purposes.
(Copyright © 2021 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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