TY - JOUR
T1 - Purification and characterization of bitiscetin, a novel von Willebrand factor modulator protein from Bitis arietans snake venom
AU - Hamako, Jiharu
AU - Matsui, Taei
AU - Suzuki, Masami
AU - Ito, Masayuki
AU - Makita, Kaori
AU - Fujimura, Yoshihiro
AU - Ozeki, Yasuhiro
AU - Titani, Koiti
N1 - Funding Information:
We thank Junko Watanabe for her technical assistance. We are grateful to Dr. Zaverio M. Ruggeri (Scripps Research Institute) and Takara Shuzo Co. for supplying monoclonal antibodies. We also thank Dr. Stephen Anderson for editing the manuscript. This study was supported in part by Grants-in-Aid from the Japanese Ministry of Education Culture and Science (to T. M. and K. T.), Fujita Health University (to K. T. and J. H.), the 96’ Cardiovascular Research (to Y. F.) and the Ryoichi Naito Foundation for Medical Research (to T. M.).
PY - 1996/9/4
Y1 - 1996/9/4
N2 - We have screened 20 snake venoms and purified a novel snake venom protein, named bitiscetin, from Bitis arietans venom that specifically binds to human von Willebrand factor (vWF) and induces platelet agglutination. Bitiscetin showed a heterodimeric structure composed of disulfide-linked α (16 kDa) and β (13 kDa) subunits on SDS-PAGE and showed a basic nature with pI value of 9.1, in contrast to botrocetin (pI 4.6), a vWF modulator isolated from another snake (Bothrops jararaca) venom. Bitiscetin-induced platelet agglutination was dependent on vWF and platelet membrane glycoprotein (GP) Ib, but not on Ca2+ and GPIIb/IIIa. vWF bound to bitiscetin but not to botrocetin electroblotted to a PVDF membrane after SDS-PAGE and this binding was diminished after reduction of disulfide bonds of bitiscetin. Bitiscetin did not cross-react to anti-botrocetin monoclonal antibodies. These results suggest that bitiscetin directly interacts with vWF and requires the protein conformation for its interaction as well as botrocetin, but its interaction manner with vWF appears to be different from that of botrocetin.
AB - We have screened 20 snake venoms and purified a novel snake venom protein, named bitiscetin, from Bitis arietans venom that specifically binds to human von Willebrand factor (vWF) and induces platelet agglutination. Bitiscetin showed a heterodimeric structure composed of disulfide-linked α (16 kDa) and β (13 kDa) subunits on SDS-PAGE and showed a basic nature with pI value of 9.1, in contrast to botrocetin (pI 4.6), a vWF modulator isolated from another snake (Bothrops jararaca) venom. Bitiscetin-induced platelet agglutination was dependent on vWF and platelet membrane glycoprotein (GP) Ib, but not on Ca2+ and GPIIb/IIIa. vWF bound to bitiscetin but not to botrocetin electroblotted to a PVDF membrane after SDS-PAGE and this binding was diminished after reduction of disulfide bonds of bitiscetin. Bitiscetin did not cross-react to anti-botrocetin monoclonal antibodies. These results suggest that bitiscetin directly interacts with vWF and requires the protein conformation for its interaction as well as botrocetin, but its interaction manner with vWF appears to be different from that of botrocetin.
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U2 - 10.1006/bbrc.1996.1345
DO - 10.1006/bbrc.1996.1345
M3 - Article
C2 - 8806626
AN - SCOPUS:0030568869
SN - 0006-291X
VL - 226
SP - 273
EP - 279
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -