湖南产尖吻蝮蛇蛇毒出血毒素性质和溶液构象的研究

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	湖南产尖吻蝮蛇蛇毒出血毒素性质和溶液构象的研究
	冉永禄; 陶勇; 叶文娟
刊名	动物学研究
	1994
卷号	15 期号:增刊页码:173-181
关键词	尖吻蝮蛇出血毒素溶液构象圆二色性
其他题名	Biochemical properties and conformation in liquid on two hemorrhagic toxins from the venom of deinagkistrodon acutus from human
合作状况	其它
中文摘要	经sephadex G-75凝胶过滤、QAE-Sephadex A-50和CM-Sephadex C-25离子交换步骤,从湖南产尖吻蝮（Deinagkistrodon acutus）蛇毒中纯化出两个出血毒素,被命名为HaHT-1和HaHT-2。用聚丙烯酰胺凝胶电泳（PAGE）和SDS-聚丙烯酰胺凝胶电泳鉴定呈单一的蛋白染色带。两个出血毒素的分子量相同,均为 23.5kDa。等电聚焦电泳测定它们的等电点分别为5.6和5.2。HaHT-1和HaHT-2均具有较强的出血活性（MHD分别为0.5和0.8μg）,都有酪蛋白水解活力,无精氨酸酯酶、胆碱酯酶和磷脂酶A2活力。用CD谱测定HaHT-l和HaHT-2的溶液构象,HaHT-1的α-螺旋、β-折叠和无规卷曲分别为36.9％、35.5％和27.6％,而HaHT-2的α-螺旋、β-折叠和无规卷曲分别为23.4％、31.3％和45.3％。pH的变化对它们构象有影响,在pH2-11范围内,酸性比碱性大。随着酸性或碱性的增加其α-螺旋含量减少,无规卷曲增加,β-折叠结构变化不大。
英文摘要	Two hemorrhagic toxins(HaHT-1 and HaHT-2) from the venom of Deinagkistrodon acutus from Hunan have been purified using Sephadex G-75 gel filtration, QAE-Sephadex A-50 and CM-Sephadex C-25 ion exchange chromatography. They were homogeneous judged by polyacrylamide gel electrophoresis and SDS-polyacrylamide electrophoresis. The both consist of single polypeptide chain with similar molecular weight of 23.5kDa. HaHT-1 is composed of 213 amino acid residues and HaHT-2 consists of 207 amino acid residues, respectively. As determined by polyacrylamide gel isoelectric focusing, their isoelectric points are 5.6 for HaHT-1 and 5.2 for HaHT-2, respectively. They were strong hemorrhagic toxins with proteolytic activity. EDTA and cysteine could inhibit both hemorrhagic and proteolytic activity but trypsin had no effect, suggesting that the toxins are metalloproteins and disulfide bridges which had contribution to the consormation of activities. Analyses for metals indicated that zinc and calcium were present in both toxins with roughly 0.5 mol zinc and 1 mol calcium per 1 mol protein. Circular dichroism (CD) was used to examine the conformation of HaHT-1 and HaHT-2. In the far CD spectrum, it was showed a wide negative peak at 217 nm for HAHT-1 and a negative peak at 2.8 nm with a shoulder peak at 217 nm for HaHT-2. Although there is some indication of a high β-sheet content, it is possible that the aromatic region contributed to this complex spectrum for HaHT-2. The secondary structure contents calculated by the method of Chen and Yang showed that HaHT-1 was found to be 36.9%α-helix, 35.5%β-sheet% and 27.6% randon corl and HaHT-2 to be 23.4%α-helix, 31.3%β-sheet% and 45.3% randon coil. The effects of pH, temperature and EDTA on CD spectra of the hemorrhagins could be found that the conformation of HaHT-1 and HaHT-2 have some changes. As they were calculated by the method of Chen and Yang, the contents of α-helix were decreased while that randon coil were increased.
收录类别	其他
语种	中文
公开日期	2010-08-10
内容类型	期刊论文
源URL	[http://159.226.149.42:8088/handle/152453/1243]
专题	昆明动物研究所_其他
作者单位	中国科学院昆明动物研究所,昆明650223
推荐引用方式 GB/T 7714	冉永禄,陶勇,叶文娟. 湖南产尖吻蝮蛇蛇毒出血毒素性质和溶液构象的研究[J]. 动物学研究,1994,15(增刊):173-181.
APA	冉永禄,陶勇,&叶文娟.(1994).湖南产尖吻蝮蛇蛇毒出血毒素性质和溶液构象的研究.动物学研究,15(增刊),173-181.
MLA	冉永禄,et al."湖南产尖吻蝮蛇蛇毒出血毒素性质和溶液构象的研究".动物学研究 15.增刊(1994):173-181.