CAJ | 학술논문

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.
목적 순화대량척신경감각섬유33.1kDa특이단백(SSP-33.1),병분석기이급결구급기신경영양활성.방법 채용쌍향전영기술비교료대서척수배배근신경섬유화복근신경섬유중단백질표체차이,용DEAE-Sephacel음리자교환층석화고효액상응효과려적방법순화료기중일충단백질,병통과원이색보초보분석료해단백질적이급결구;우통과경전적신경영양활성적모형-계배배근절(DRG)체외배양모형,측정해단백적생물학활성.결과 순화적단백질분자량위33.1,등전점위5.52,기이급결구중α라선점20.8%,β편층점54.8%,전각점7.3%,무규권곡점17.1%.체외실험표명해단백질능촉진계배배근경절돌기적생장.결론 순화료대서감각신경33.1kDa특이단백,해단백질이급결구이β편층위주,체외실험표명해단백질대체외배양적계배배신경절유신경영양활성.
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.