Please use this identifier to cite or link to this item: https://repositorio.unifesp.br/handle/11600/36249
Title: A Novel Hyaluronidase from Brown Spider (Loxosceles intermedia) Venom (Dietrich's Hyaluronidase): From Cloning to Functional Characterization
Authors: Ferrer, Valeria Pereira
Mari, Thiago Lopes de
Gremski, Luiza Helena
Silva, Dilza Trevisan
Silveira, Rafael Bertoni da
Gremski, Waldemiro
Chaim, Olga Meiri
Senff-Ribeiro, Andrea
Nader, Helena Bonciani [UNIFESP]
Veiga, Silvio Sanches
Univ Fed Parana
Univ Estadual Ponta Grossa
Catholic Univ Parana
Universidade Federal de São Paulo (UNIFESP)
Issue Date: 1-May-2013
Publisher: Public Library Science
Citation: Plos Neglected Tropical Diseases. San Francisco: Public Library Science, v. 7, n. 5, 12 p., 2013.
Abstract: Loxoscelism is the designation given to clinical symptoms evoked by Loxosceles spider's bites. Clinical manifestations include skin necrosis with gravitational spreading and systemic disturbs. the venom contains several enzymatic toxins. Herein, we describe the cloning, expression, refolding and biological evaluation of a novel brown spider protein characterized as a hyaluronidase. Employing a venom gland cDNA library, we cloned a hyaluronidase (1200 bp cDNA) that encodes for a signal peptide and a mature protein. Amino acid alignment revealed a structural relationship with members of hyaluronidase family, such as scorpion and snake species. Recombinant hyaluronidase was expressed as N-terminal His-tag fusion protein (similar to 45 kDa) in inclusion bodies and activity was achieved using refolding. Immunoblot analysis showed that antibodies that recognize the recombinant protein cross-reacted with hyaluronidase from whole venom as well as an anti-venom serum reacted with recombinant protein. Recombinant hyaluronidase was able to degrade purified hyaluronic acid (HA) and chondroitin sulfate (CS), while dermatan sulfate (DS) and heparan sulfate (HS) were not affected. Zymograph experiments resulted in similar to 45 kDa lytic zones in hyaluronic acid (HA) and chondroitin sulfate (CS) substrates. Through in vivo experiments of dermonecrosis using rabbit skin, the recombinant hyaluronidase was shown to increase the dermonecrotic effect produced by recombinant dermonecrotic toxin from L. intermedia venom (LiRecDT1). These data support the hypothesis that hyaluronidase is a spreading factor. Recombinant hyaluronidase provides a useful tool for biotechnological ends. We propose the name Dietrich's Hyaluronidase for this enzyme, in honor of Professor Carl Peter von Dietrich, who dedicated his life to studying proteoglycans and glycosaminoglycans.
URI: http://repositorio.unifesp.br/handle/11600/36249
ISSN: 1935-2735
Other Identifiers: http://dx.doi.org/10.1371/journal.pntd.0002206
Appears in Collections:Em verificação - Geral

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