摘要
从暴露于污染物蒽的鲤肝脏中提取出芳香烃受体,并采用紫外可见分光光度计进行最大吸收波长扫描,再通过离子交换层析法分离纯化鲤体内的芳香烃受体,并采用聚丙烯酰胺凝胶电泳对受体进行鉴定,最后通过傅里叶变换红外光谱探讨芳香烃受体与蒽结合后的结构变化。结果表明,提取的芳香烃受体在280 nm处有特征吸收峰,测得质量浓度为27.3 mg/mL。通过离子交换层析法纯化了单一组分的受体蛋白质,其相对分子质量约为110 k D。测定了芳香烃受体的红外光谱,与蒽结合后其在一定波段的谱峰发生了明显位移,表明污染物配体蒽能与芳香烃受体结合,并对其蛋白二级结构产生明显影响。
This paper is aimed to gain the aryl hydrocarbon receptor（AHR）from the organisms of Cyprinus carpio as well as to pursue the effects of the polycyclic aromatic hydrocarbons（PAHs）on the said hydrocarbon receptor.To achieve the corresponding research goals,we have tried to take out the AHR from the liver of the aforementioned fish to expose it to anthracene of10 mg/L via the ultraviolet-visible spectrophotometer（UV-Vis）by scanning the maximum absorption wavelength of the extracts and determinating the content concentrations of AHR in the said extracts with the method of Bradford with UV-Vis.And,then,the purified extracts and AHR gained with high purity by means of the ion exchange chromatography（IEC）.Further studies have been done to identify AHR through the sodium dodecyl sulfatepolyacrylamide gel electrophoresis（SDS-PAGE）.All having been done,a detailed discussion has been done with the structural changes of AHR after the interaction with anthracene via the Fourier Transform Infrared（FTIR）.The results we have achieved from the above experiments verify that it is possible for the extraction of AHR to reach its characteristic absorption peak to a degree of 280nm,known as the protein absorption feature,with the standard absorption curve of protein being Y=0.0067X＋0.0238 and R2is 0.995 8.Then,the extracted concentration of AHR has been found equal to 27.3 mg/m L which has been worked out via the equation.On the other hand,the SDS-PAGE result can be regarded to attain a satisfactory level for purification with a single component of AHR gained through the method of IEC.Accurately speaking,the molecular weight of AHR should be taken as about 110 k D.What is more,the FTIR experimental results prove that the AHR shift can be said to reach its peak value in a wavelength range among amide bondⅠ,amide bondⅢand the fingerprint region,whereas the shift to lower wavelengths implies a decrease in the amide bonds.Thus,it can be seen that the anthracene ligand is in a position to combine and interact with AHR,which has a significant effect on the protein secondary structure.Therefore,it is entitled to have a significant impact on the transcription and expression of the receptor in the organism and in turn have a negative impact on the maintenance of the dynamic equilibrium,growth,reproduction and biological behaviors.
出处
《安全与环境学报》
CAS
CSCD
北大核心
2017年第3期1205-1209,共5页
Journal of Safety and Environment
基金
上海市自然科学基金项目(15ZR1401500)
关键词
环境科学技术基础学科
蒽
芳香烃受体
离子交换层析
傅里叶变换红外光谱
basic disciplines of environmental science and technology
anthracene
aryl hydrocarbon receptor
ionexchange column chromatography
Fourier transform infrared spectroscopy
