大疣蛛毒素-VI的分离纯化及部分生物学活性鉴定

Purification,Characterization of Raventoxin-VI from the Venom of the Spider Macrothele raveni

从雷氏大疣蛛(Macrotheleraveni)粗毒中,结合阳离子交换和反相高效液相色谱分离到一种多肽神经毒素,命名为大疣蛛毒素 VI(Raventoxin VI).经MALDI TOF质谱分析,它的分子量为(5371.6±0.5)Da.利用Edman降解气相蛋白质测序仪测得其氨基酸序列是NH2 NIIKGRVVKLCGGCAQKCCDREPRCDPCRTCVENVGT GGGYLSSNKKCNGS COOH,其中8个Cys形成4对二硫键.Raventoxin Ⅵ能阻断小鼠膈神经膈肌标本神经肌肉接头传递,脑室注射能使小鼠瘫软.从一级结构上没有发现与该毒素有较高相似性的其他蜘蛛毒素.

A novel neurotoxin named RaventoxinVI is purified from the venom of the spider Macrothele raveni by ion exchange chromatography and reverse phase high performance liquid chromatography.The molecular weight of RaventoxinVI is (5 371.6±0.5) Da,identified by MALDITOF mass spectrometry,which is in good agreement with the complete amino acid sequence of NH2NIIKGRVVKLCGGCAQKCCDREPRCDPCRTCVENVGTGGGYLSSNKKNCGSCOOH determined by automatic Edman degradation.RaventoxinVI is a novel neurotoxin which can paralyze rat after intracerebroventricular injection and block the neuromuscular transmission of isolated rat phrenic nerve diaphragm preparation. The 51residues peptide,which contained 8 Cys and formed four disulfide bonds,shows no homology with other spider toxins.