TY - JOUR
T1 - Purification and characterization of a new weak hemorrhagic metalloproteinase BmHF-1 from Bothrops marajoensis snake venom
AU - Torres-Huaco, Frank Denis
AU - Ponce-Soto, Luis Alberto
AU - Martins-De-Souza, Daniel
AU - Marangoni, Sergio
N1 - Funding Information:
Acknowledgments The authors thank Mr. Paulo A. Baldasso for technical assistance. This work was supported by the National Council for Scientific and Technological Development (CNPq) and FAPESP process 06/51566-0. It is part of Frank Denis Torres-Huaco Mg thesis.
PY - 2010/8
Y1 - 2010/8
N2 - 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.
AB - 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.
KW - BmHF-1
KW - Bothrops marajoensis
KW - HPLC-RP
KW - Hemorrhagic activity
KW - Snake venom metaloproteases
UR - http://www.scopus.com/inward/record.url?scp=77956265429&partnerID=8YFLogxK
U2 - 10.1007/s10930-010-9267-z
DO - 10.1007/s10930-010-9267-z
M3 - Article
C2 - 20607373
AN - SCOPUS:77956265429
SN - 1572-3887
VL - 29
SP - 407
EP - 416
JO - Protein Journal
JF - Protein Journal
IS - 6
ER -