Department of Biochemistry, Manasagangotri, University of Mysore, Mysore 570 006, India.
Toxicon. 2011 Feb;57(2):248-58. doi: 10.1016/j.toxicon.2010.11.018. Epub 2010 Dec 13.
The current study describes the biochemical, biophysical and pharmacological properties of Hag-protease-II from Hippasa agelenoides spider venom gland extract. The Hag-protease-II was purified to homogeneity using gel filtration and ion-exchange chromatography. The molecular mass was found to be 28.749 kDa by MALDI-TOF mass spectrometry. PMSF abolished the activity while EDTA, EGTA, IAA and 1, 10-phenanthrolene did not. Hag-protease-II hydrolyzed casein, fibrinogen and fibrin, however it did not hydrolyze gelatin, fibronectin and collagen types- I and IV. It was non-lethal and devoid of hemorrhagic, myotoxic and edema forming activities. It dose dependently reduced re-calcification time of citrated human plasma. Strikingly; the Hag-protease-II coagulated the factor X deficient congenital human plasma. It hydrolyzed Bβ-chain but, did not degrade Aα- and γ-chains of fibrinogen while, it hydrolyzed α-polymer and α-chain but not the β-chain and γ-γ dimers of partially cross-linked fibrin clot. The Hag-protease-II induced aggregation of human platelets in PRP dose dependently, however it did not interfere in collagen induced aggregation of PRP and washed human platelets.
本研究描述了 Hippasa agelenoides 蜘蛛毒液腺提取物中 Hag 蛋白酶-II 的生化、生物物理和药理学特性。Hag 蛋白酶-II 通过凝胶过滤和离子交换层析纯化至均一性。MALDI-TOF 质谱法发现其分子量为 28.749 kDa。PMSF 使其失活，而 EDTA、EGTA、IAA 和 1,10-菲咯啉则没有。Hag 蛋白酶-II 水解酪蛋白、纤维蛋白原和纤维蛋白，但不水解明胶、纤维连接蛋白和胶原 I 型和 IV 型。它无细胞毒性，无出血、肌毒性和水肿形成活性。它剂量依赖性地缩短了柠檬酸化人血浆的再钙化时间。引人注目的是，Hag 蛋白酶-II 凝固了因子 X 缺乏的先天性人血浆。它水解 Bβ 链，但不降解纤维蛋白原的 Aα-和 γ-链，而水解部分交联纤维蛋白凝块的 α-聚合物和 α-链，但不水解 β-链和 γ-γ 二聚体。Hag 蛋白酶-II 依赖性地诱导 PRP 中的人血小板聚集，但不干扰胶原诱导的 PRP 和洗涤人血小板的聚集。
