Potent histamine-releasing activity of atrahagin, a novel snake venom metalloproteinase.
Poisonous snakebite wound is a popular disease worldwide. However, the pathogenesis remains unclear. In the present study, a novel metalloproteinase atrahagin in Chinese cobra (Naja atra) snake venom was purified, using heparin-sepharose followed by Superdex 75 gel filtration chromatography. Apart from its alpha-fibrinogenase activity, atrahagin potently activated human colon, lung and tonsil mast cells with the net histamine release being 25.9+/-4.4, 17.0+/-1.9, 13.2+/-3.6%, respectively. Time course studies revealed that the peak histamine release induced by atrahagin occurred at 12, 12 and 8 min following incubation of the enzyme with colon, lung and tonsil mast cells, respectively. The response of mast cells to atrahagin was abolished by preincubation of the cells with metabolic inhibitors or pertussis toxin, and by removal of Ca2+ and Mg2+ from the challenge buffer. In conclusion, activation of human mast cells by atrahagin indicated that the enzyme might contribute to the pathogenesis of snakebite wound.
Wei
Ji-Fu
JF
Allergy and Inflammation Research Institute, The Shantou University Medical College, Shantou, Guangdong Province 515041, China. weijifu@hotmail.com
Mo
Ya-Zhen
YZ
Qiao
Li-Ya
LY
Wei
Xiao-Long
XL
Chen
Han-Qiu
HQ
Xie
Hua
H
Fu
Yi-Ling
YL
Wang
Wan-Yu
WY
Xiong
Yu-Liang
YL
He
Shao-Heng
SH
D000818:Animals; D002478:Cells, Cultured; D004546:Elapid Venoms / Q000737:chemistry* / Q000494:pharmacology; D005340:Fibrinogen / Q000378:metabolism; D006636:Histamine Release / Q000187:drug effects*; D006801:Humans; D008407:Mast Cells / Q000166:cytology / Q000378:metabolism*; D045726:Metalloproteases / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D030162:Reptilian Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D013997:Time Factors
D004546:Elapid Venoms; D030162:Reptilian Proteins; D005340:Fibrinogen; D045726:Metalloproteases; C511413:atrahagin