Purification and characterization of a new weak hemorrhagic metalloproteinase BmHF-1 from Bothrops marajoensis snake venom

Frank Denis Torres-Huaco, Luis Alberto Ponce-Soto, Daniel Martins-De-Souza, Sergio Marangoni

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12 Scopus citations

Abstract

BmHF-1, from the venom of Bothrops marajoensis, was purified by Sephadex G-75 and HPLC-RP on μ-Bondapak C-18 column chromatography. It presented a molecular mass of 27162.36 Da determined by MALDI-TOF MS. BmHF-1 had a sequence of 238 residues of amino acids. The multiple alignment of its amino acid sequence and those of other snake venom metalloproteinases showed high structural similarity, mainly among P-I class. The enzyme initially cleaves the Aα-chain of fibrinogen, followed by the Bβ-chain, and shows no effects on the γ-chain. BmHF-1 had, caseinolytic and weakly hemorrhagic activities, which were inhibited by EDTA. In contrast, PMSF did not affect these activities. The caseinolytic activity of BmHF-1 had a pH optimum of 8.0 and was stable in solution up to 40 °C; activity was completely lost at ≥70 °C. The proteolytic activity was also inhibited by sDa (opossum sera) and Da2-1, Da2-II, antihemorrhagic factors isolated from the opossum sera of Didelphis albiventris. BmHF-1 presents weak hemorrhagic activity, with a MHD of 41.14 μg and it induces dose-dependent edema. We could concluded that, despite its weak hemorrhagic activity, BmHF-1 contributes to local tissue damage by inducing edema, releasing pharmacologically active mediators from protein precursors due to its enzymatic action.

Original languageEnglish
Pages (from-to)407-416
Number of pages10
JournalProtein Journal
Volume29
Issue number6
DOIs
StatePublished - Aug 2010
Externally publishedYes

Keywords

  • BmHF-1
  • Bothrops marajoensis
  • HPLC-RP
  • Hemorrhagic activity
  • Snake venom metaloproteases

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