Purification and Characterization of a New Weak Hemorrhagic Metalloproteinase BmHF-1 from Bothrops marajoensis Snake Venom

Author(s)

Torres-Huaco, Frank Denis ^[1] ; Ponce-Soto, Luis Alberto ^{[1, 2]} ; Martins-de-Souza, Daniel ^{[3, 1]} ; Marangoni, Sergio ^[1]

Affiliation

^[1] Univ Estadual Campinas, Dept Biochem, Inst Biol, BR-13083970 Campinas, SP - Brazil

^[2] Univ Estadual Campinas, Dept Pharmacol, Fac Med Sci FCM, BR-13083970 Campinas, SP - Brazil

^[3] Max Planck Inst Psychiat, D-80804 Munich - Germany

• Document type: Journal article
• Source: The Protein Journal; v. 29, n. 6, p. 407-416, AUG 2010.
• Web of Science Citations: 8

Abstract

BmHF-1, from the venom of Bothrops marajoensis, was purified by Sephadex G-75 and HPLC-RP on mu-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 A alpha-chain of fibrinogen, followed by the B beta-chain, and shows no effects on the gamma-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 A degrees C; activity was completely lost at a parts per thousand yen70 A degrees 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 mu g 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. (AU)