虎纹捕鸟蛛毒素-Ⅰ天然突变体的分离纯化与鉴定  

Purification and Identification of a Natural Mutant of Huwentoxin-Ⅰ from the Venom of Ornithoctonus huwena

作　　者：何小珍[1] 胡卫军[1] 曹税[1] 王贤纯[1] 梁宋平[1] 

机构地区：[1]湖南师范大学生命科学学院蛋白质化学与发育生物学教育部重点实验室,中国长沙410081

出　　处：《湖南师范大学自然科学学报》2005年第3期76-79,共4页Journal of Natural Science of Hunan Normal University

基　　金：国家自然科学基金资助项目(30370260)

摘　　要：通过离子交换及反相HPLC,从虎纹捕鸟蛛粗毒中分离得到一种虎纹毒素-Ⅰ天然突变体,MALDI-TOF质谱仪分析表明该多肽天然突变体分子量为3 636 Da.Edman测序结合串联质谱和氨基酸组成分析鉴定出该天然突变体的序列为H2N-ACKGV FDACT PGKNE CCPNR VCSDK HKWCK WQ-COOH.其中6个Cys形成3对二硫键.小鼠脑室注射实验表明,该天然突变体能使小鼠致死,但不能阻断小鼠膈神经-膈肌标本的神经肌肉接头传递.本研究结果表明,K32残基对于HWTX-Ⅰ的生物学活性有关键性作用.A natural mutant of Huwentoxin- Ⅰ was purified and identified from the venom of the spider Ornithotonus huwena by ion exchange chromatography and reversed phase HPLC. The molecular weight of the mutant is 3636 Da determined by MALDI-TOF mass spectrometry and the amino acid sequence is H2N-ACKGV FDACT PGKNE CCPNR VCSDK HKWCK WQ-COOH determined by Edman degradation acid composition analysis. The six Cys residues in the mutant fo in combination with tandem mass spectrometry and amino rm three pairs of disulfide bridge. The natural mutant can cause the death of adult mouse after intracerebroventricular injection but can not block the neuromuscular transmission of the isolated mouse phrenie nerve-diaphragm preparation. This result indicates that the residue K32 is one of the key residues related to the biological activity of HWTX- Ⅰ .

关 键 词：天然突变体 虎纹捕鸟蛛毒素-Ⅰ 虎纹捕鸟蛛 

分 类 号：Q51[生物学—生物化学]