龜殼花蛇毒中出血毒素之分離及其物理化學特性

Purification and Characterization of Hemorrhagic Toxins from the Venom of Trimeresurus Mucrosquamatus

郭國華(Kou-Wha Kuo) ； 張均昌(Chun-Chang Chang)

《The Kaohsiung Journal of Medical Sciences》 2卷12期 (1986/12) Pp. 722-731

https://doi.org/10.6452/KJMS.198612.0722

摘要

龜殼花(Trimeresurus mucrosquamatus)蛇毒以 DEAE-Sephadex A-50、DEAE-Sephacel 以及 DEAE-Cellulose 等之陰離子交換層析法配合膠過濾法純化得到三種含有鈣、鎂及鋅等金屬之出血毒蛋白；MH-Ⅱa、MH-Ⅱb 及 MH-Ⅲ。其分子量分別為 117,000、117,000 及 22,000，pI為4.8、5.4 及 5.7 之酸性蛋白質。它們皆具有酯解酶、蛋白質分解酶以及磷脂酶A2 之活性，其中酯解酶活性以MH-Ⅱa 為最高，同時其出血活性亦最強，兩者呈平行關係，顯示酯解酶活性可能與出血活性有關。此外MH-Ⅱa 亦具有相當高的蛋白質分解酶活性，但與出血活性並不成平行關係。這此出血毒蛋白之出血活性會受不同濃度的 EDTA 所抑制，其中以 MH-Ⅲ為最明顯，添加 EDTA 後其螢光強度增加40%，表示金屬離子為維持其活性構造所必需。這些出血毒蛋白並無溶血作用，但在phosphatidylcholine 的存在下却具有溶血作用，顯示它們具有 phospholipase A2 的活性。分離出來之三種出血毒蛋白均為醣蛋白，其中以 MH-Ⅱb 含30% 六碳醣類為最高。在免疫學上，它們均具有不同的抗原性。

並列摘要

Three hemorrhagic toxins designated as MH-IIa, Mh-IIb and MH-III were purified from the venom of Trimeresurus mucrosquamatus (Taiwan habu) by successive column chromatography on DEAE-Sephadex A-50, DEAE-Sephacel and DEAE-Cellulose (DE-52), then followed by gel filtration on Sephadex G-100 or G-150. These purified toxins proved to be homogeneous by polyacrylamide gel electrophoresis and immunodiffusion. Analysis indicated that these toxins are acidic glycoproteins Containing Ca2+ , Mg2+ and Zn2+ with molecular weights at about 117,000, 117,000 and 22,000, and with pI values of 4.8, 5.4 and 5.7, respectively. The toxins studied possessed protease, arginineester hydrolase and phospholipase A2 activities. Higher proteolytic activity was found in MH-IIa which had the most potent hemorrhagic activity among them. The hemorrhagic activity of these toxins was completely blocked by EDTA, indicating that one or more of these metal ions are essential for hemorrhagic activity. On immunodiffusion, they were immunologically unidentical.

並列關鍵字

hemorrhagic toxins ； arginine esterase and protease