赤尾鮐蛇毒中出血毒素之精製及生化與免疫學特性

Purification And Characterization Of Hemorrhagic Toxins From The Venom Of Trimeresurus Gramineus

陳玲娜(Lyn-Nou CHEN) ； 張均昌(Chun-Chang CHANG)

《The Kaohsiung Journal of Medical Sciences》 1卷2期 (1985/02) Pp. 113-122

https://doi.org/10.6452/KJMS.198502.0113

摘要

赤尾鮐(Trimeresurus gramineus)蛇毒以DEAE-Cell-ulose與DEAE-Sephacel之陰離交換層析法以及膠過濾法純化得到三種含有鈣、鋅及鎂等金屬之酸性出血毒蛋白；GH-IIIa、GH-IIIb及GH-IV。其分子量分別為77,000、68,000及15,000, pI為5.8、5.6及4.6。這些毒蛋白皆具有蛋白質分解醉活性，此外，GH-IIIa和IIIb 有醋解梅活性，GH-IV有磷脂醇A2活性，GH-IIIa及IV有纖維蛋白原分解活性， GH-IIIb則具有類凝血醇活性。抗GH-IIIb血清及其Fab對GH-IIIa與IIIb的出血及蛋白質分解醃兩種活性有不同程度的中和作用，顯示蛋白質分解醃活性可能是引起出血的重要因素之一，但出血活性與蚤白質分解醉活性的作用部位可能相異。GH-IIIa與IIIb在免疫學上具有類似的抗原結構，但與GH-IV則為相異的抗原性。由測定GH-IIIb與Fab反應所形成之可溶性複合物的分子量結果，得知其分子至少具有三個抗體決定基。

並列摘要

Trimeresurus gramineus (Green habu) venom was separated into 18 fractions by chromatography on DEAE-Cellulose. Among the four fractions having hemorrhagic activity, fractions 12 and 16 , designated as GH-III and GH-IV, were further purified on DEAE-Sephacel followed by gel filtration on Sephadex G-100 or G-75. GH-III was further separated into two hemorrhagic toxic pro-teins, GH-III and IIIb. The purified toxins were proved to be homogeneous by polyacrylamide gel electrophoresis, and single poly-peptide containing carbohydrates and metals, Zn2+ , Ca2+ and Mg2+. The molecular weight of GH-III, IIIb and IV were determined to be about 77,000, 68,000 and 15,000 and pI values, 5.8 ,5.6 and 4.6, respectively.
Three hemorrhagic toxins possessed proteolytic activity. Potent arginine ester hydrolase was found in GH-IIIa and IIIb, while phospholipase A2 activity was found in GH -IV. In addition , GH-IIIa and IV hada weak fibrinogenolytic activity, and GH- IIIb possessed thrombin-like enzyme activity.
On immunodiffusion, GH-IIIb showeda partial immunologic identity with GH-IIIa， but was non-identical with GH-IV. Although the hemorrhagic activity of both GH-IIIa and IIIb was completely neutralized by anti-GH-IIIb serum, their proteolytic activity was only partially neutralized. These results indicate that the active site (s) of hemorrhagic activity may be different from that of proteolytic activity. The hemorrhagic activity of GH-IIIb was completely neutralized by Fab while that of GH –IIIa was partially inactivated. The proteolytic activity of both GH-IIIa and IIIb, however, was not neutralized by Fab. An estimate of the molecular weight of the soluble complex formed from Fab with GH-IIIb indicates that at least three antigenic sites in the toxin molecule can be simultaneously occupied by the antibody.

並列關鍵字

hemorrhagic toxins ； anti-hemorrhagic toxin antibody