Jiang M, Häggblad J, Heilbronn E, Rydqvist B, Eaker D
Unit of Neurochemistry and Neurotoxicology, University of Stockholm, Sweden.
Toxicon. 1987;25(7):785-92. doi: 10.1016/0041-0101(87)90129-2.
A toxic component (AgTx) from the venom of Agkistrodon halys (Pallas) was isolated using DEAE-cellulose DE11 and CM-Sephadex C50 column chromatography and finally purified to homogeneity by FPLC on a MonoQ column. The toxin is a neutral (pI 6.9) single chain polypeptide with a mol. wt of 14,000 and an amino acid composition (123 residues) roughly similar to that of notexin. AgTx was found to have phospholipase A2 activity which was dependent on calcium and stimulated by sodium deoxycholate. The toxin caused efflux of 2-deoxy-(1-3H)-glucose-6-phosphate (a cell membrane integrity probe) as well as of [3H]acetylcholine from rat brain synaptosomes. No cell membrane damage was induced by AgTx on cultured N1E 115 neuroblastoma cells and chick myotube cultures. The LD50 ws 150 micrograms/kg (i.p.) in mice. The main symptom observed was respiratory paralysis. The results obtained show that AgTx can be classified as a toxic phospholipase A2 with a presynaptic site of action.
利用DEAE - 纤维素DE11和CM - 葡聚糖凝胶C50柱色谱法从蝮蛇(Agkistrodon halys (Pallas))毒液中分离出一种毒性成分(AgTx),最后通过在MonoQ柱上进行快速蛋白质液相色谱(FPLC)将其纯化至均一。该毒素是一种中性(pI 6.9)单链多肽,分子量为14,000,氨基酸组成(123个残基)与诺毒素大致相似。发现AgTx具有依赖于钙且受脱氧胆酸钠刺激的磷脂酶A2活性。该毒素导致大鼠脑突触体中2 - 脱氧 -(1 - 3H)- 葡萄糖 - 6 - 磷酸(一种细胞膜完整性探针)以及[3H]乙酰胆碱外流。AgTx对培养的N1E 115神经母细胞瘤细胞和鸡肌管培养物未诱导细胞膜损伤。在小鼠中,腹腔注射的半数致死量(LD50)为150微克/千克。观察到的主要症状是呼吸麻痹。所得结果表明,AgTx可归类为一种具有突触前作用位点的毒性磷脂酶A2。