[利用圆二色光谱分析大肠杆菌中β2微球蛋白的二级结构]
[Analyzing secondary structure of beta2 microglobulin in Escherichia coli with circular dichroism spectrum].
作者信息
Gao Fengshan, Xia Chun, Zhang Qiang, Li Yungang, Li Xinsheng, Xu Chongbo, Bai Jing
机构信息
Dalian University, Bioengineering College, Dalian 116622, China.
出版信息
Wei Sheng Wu Xue Bao. 2009 Dec;49(12):1596-600.
OBJECTIVE
In order to study the structure and function of beta2 microglobulin (beta2 m).
METHODS
We sub-cloned the mature peptide of beta2 m into the p2X plasmid and transformed them to Escherichia coli TB1. The recombinant bacterium was induced to be expressed and the expressed fusion protein was detected by SDS-PAGE and western blot. After purifying and cleaving with Factor Xa, we separated the monomer protein beta2 m from MBP. Finally, we determined the secondary structure of the beta2 m protein by circular dichroism (CD) spectrum.
RESULTS
MBP-beta2 m was 52.1 kDa, and the monomer protein beta2 m was 10.6 kDa. The alpha-helix, beta-sheet, turn, and random coil of the fusion protein showed 0, 45, 8 and 45 amino acids, respectively, detected by CD estimation.
CONCLUSION
The beta2 m protein had correct secondary structure and could be used for further research of peptide binding in vitro.
目的
为研究β2微球蛋白（β2m）的结构与功能。
方法
我们将β2m的成熟肽亚克隆至p2X质粒，并将其转化至大肠杆菌TB1。诱导重组菌表达，通过SDS-PAGE和蛋白质免疫印迹法检测表达的融合蛋白。经纯化并用凝血因子Xa切割后，我们从麦芽糖结合蛋白（MBP）中分离出单体蛋白β2m。最后，我们通过圆二色光谱（CD）测定β2m蛋白的二级结构。
结果
MBP-β2m为52.1 kDa，单体蛋白β2m为10.6 kDa。通过CD估算，融合蛋白的α螺旋、β折叠、转角和无规卷曲分别显示为0、45、8和45个氨基酸。
结论
β2m蛋白具有正确的二级结构，可用于进一步的体外肽结合研究。
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