感觉神经33.1 kDa特异蛋白二级结构及其神经营养活性探讨

感觉神经33.1 kDa特异蛋白二级结构及其神经营养活性探讨

沈建英, 俞庆声, 王琪, 李泉, 蒲小平^*

北京大学药学院分子与细胞药理学系, 100083

收稿日期:2003-02-17 修回日期:2003-05-10 出版日期:2003-06-15 发布日期:2003-06-15
通讯作者: 蒲小平*

Secondary Structure and Neurotrophic Effect of a 33.1 kDa Specific Protein (SSP-33.1) in Spinal Sensory Ganglia

SHEN Jian-ying, YU Qing-sheng, WANG Qi, LI Quan, PU Xiao-ping^*

Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Sciences, Peking University, Beijing 100083, China

Received:2003-02-17 Revised:2003-05-10 Online:2003-06-15 Published:2003-06-15
Contact: PU Xiao-ping*

摘要/Abstract

摘要： 目的纯化大量脊神经感觉纤维33.1kDa特异蛋白(SSP-33.1), 并分析其二级结构及其神经营养活性. 方法采用双向电泳技术比较了大鼠脊髓背背根神经纤维和腹根神经纤维中蛋白质表达差异, 用DEAE-Sephacel阴离子交换层析和高效液相凝胶过滤的方法纯化了其中一种蛋白质, 并通过圆二色谱初步分析了该蛋白质的二级结构; 又通过经典的神经营养活性的模型-鸡胚背根节(DRG)体外培养模型, 测定该蛋白的生物学活性. 结果纯化的蛋白质分子量为33.1, 等电点为5.52, 其二级结构中α螺旋占20.8%, β片层占54.8%, 转角占7.3%, 无规卷曲占17.1%.体外实验表明该蛋白质能促进鸡胚背根经节突起的生长. 结论纯化了大鼠感觉神经33.1kDa特异蛋白, 该蛋白质二级结构以β片层为主, 体外实验表明该蛋白质对体外培养的鸡胚背神经节有神经营养活性.

关键词: 感觉神经特异33.1kDa蛋白 SSP-33.1, 双向电泳 2-DE, 高效液相 HPLC, 圆二色谱 CD, 神经营养活性

Abstract: Aim To analyze the secondary structure and neurotrophic effect of a specific protein in sensory neurons. Methods Comparison of the proteins expressed in the rat spinal sensory neurons and motor neurons was made by two-dimensional electrophoresis. One specific protein in sensory neurons was isolated and purified by DEAE-Sephacel ion exchange chromatography and high performance liquid chromatography. A primary analysis of its secondary structure by circular dichroism, and its neurotrophic effects were investigated using the model of dorsal root ganglia (DRG) cultured in vitro. Results The molecular weight and isoelectric point of the protein were 33.1 kDa and 5.52, respectively. Its circular dichroism showed that there were 20.8% α-helix, 54.8% β-sheet, 7.3% turn, and 17.1% random coil in its secondary structure. Biological experiments showed that the protein could promote the neurite outgrowth of DRG. Conclusion A specific protein in spinal sensory tissue with molecular weight of 33.1 kDa has been purified. There is mainly β-sheet in the secondary structure of the protein. And the protein has neurotrophic effects in the model of DRG.

Key words: spinal sensory specific 33.1 kDa protein, spinal sensory specific 33.1 kDa protein, two dimensional electrophoresis, two dimensional electrophoresis, high performance liquid chromatography, high performance liquid chromatography, circular dichroism, circular dichroism, neurotrophic effect, neurotrophic effect

中图分类号:

R963

Supporting: Foudation item: Supported by the National Program for Preperiod Key Basic Research Projects.
Biography: Shen Jianying female , Master graduate.
^*Corresponding author. Tel.: 8610-62092648; fax: 8610-62092648

沈建英, 俞庆声, 王琪, 李泉, 蒲小平^*. 感觉神经33.1 kDa特异蛋白二级结构及其神经营养活性探讨[J]. .

SHEN Jian-ying, YU Qing-sheng, WANG Qi, LI Quan, PU Xiao-ping^*. Secondary Structure and Neurotrophic Effect of a 33.1 kDa Specific Protein (SSP-33.1) in Spinal Sensory Ganglia[J]. .