摘要
经硫酸铵分级沉淀、SP-Sepharose FF离子交换层析、Blue Sepharose CL-6B亲和层析和Superdex75分子筛层析从苦瓜籽中纯化出苦瓜核糖体失活蛋白．纯化的蛋白经IEF、梯度PAGE、SDS-PAGE和Periodate—Schiff分析均为单一蛋白着色带或单一糖蛋白着色带．根据SDS—PAGE计算其相对分子量为3．0×10^4Da，IEF显示其pI为9．0．N端测序结果为Asp—Val—Ser—Phe-Arg．采用傅里叶变换红外光谱（FTIR）和圆二色谱（CD谱）对纯化RIP的二级结构进行定量分析，结果显示其α-螺旋和β-折叠含量均在30％以上．
A RIP from the Momordica charantia seeds was purified by precipitation of 30% -70% of （NH4）2SO4 and chromatographies on SP-Sepharose FF, Blue Sepharose CL-6B and Superdex 75. The results of IEF-PAGE, Gradient-PAGE, SDS-PAGE and Periodate-Schiff staining after SDS-PAGE showed that RIP was a homogeneous single band or a glycoprotein band with pI of 9.0. The carbohydrate content of purified RIP was about 1.05%. The molecular weight of 30 kDa was determined by SDS-PAGE. The N-terminal sequence of RIP was determined to be Asp-Val-Ser-Phe-Arg. The secondary structure of RIP was also studied by CD spectra and FTIR.
出处
《四川大学学报（自然科学版）》
CAS
CSCD
北大核心
2009年第4期1203-1206,共4页
Journal of Sichuan University(Natural Science Edition)
基金
国家自然科学基金(30770232)
关键词
苦瓜籽
核糖体失活蛋白
二级结构
Momordica charantia seeds, ribosome inhibiting protein, secondary structure
