椰子花粉过敏原profilin蛋白体外重折叠过程的光谱学研究

Spectroscopic Studies on Refolding Process in Vitro of Pan-AllergenProfilin in Coco Pollen

在基因工程技术中,采用原核表达系统获得的重组蛋白通常都是以包涵体的形式存在。利用圆二色性光谱、荧光光谱和同步荧光光谱对尿素诱导的重组椰子花粉泛过敏原profilin蛋白包涵体的复性过程进行了系统的光谱学研究,得到了profilin蛋白复性过程的光谱学特征;结合生物信息学方法,通过对profilin蛋白的二级结构和三级结构的预测,进一步分析了复性过程中profilin蛋白构象变化的特点,初步建立了一种新的检测重组变应原蛋白复性程度的光谱学实验方法,为重组蛋白包涵体复性机理的深入研究进行了有益的探索。

Recombinant proteins expressed by prokaryotic expression system are normally in the form of inclusion.In the present paper,refolding process of recombinant pan-allergen profilin protein induced by urea has been investigated by using circular dichroism spectra,fluorescence spectra,synchronous fluorescence spectra systematically.And the spectral characteristics of the renaturation were obtained.In addition,bioinformatics methods including predications of secondary and tertiary structures have also been used to explain the spectral characteristics and analyze the conformational changes of the protein during renaturation in vitro.Results from this study should be useful to the establishment of a spectral method examining the extent of protein renaturation,and be helpful to the understanding of the mechanism of renaturation of recombinant protein.