MEASUREMENT of Protein 53 Diffusion Coefficient in Live HeLa Cells Using Raster Image Correlation Spectroscopy (RICS).

Journal of Biomaterials and Nanobiotechnology Pub Date : 2010-10-01 DOI:10.4236/jbnb.2010.11004

Sungmin Hong, Ying-Nai Wang, Hirohito Yamaguchi, Harinibytaraya Sreenivasappa, Chao-Kai Chou, Pei-Hsiang Tsou, Mien-Chie Hung, Jun Kameoka

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引用次数: 12

Abstract

We have applied Raster Image Correlation Spectroscopy (RICS) technique to characterize the dynamics of protein 53 (p53) in living cells before and after the treatment with DNA damaging agents. HeLa cells expressing Green Fluorescent Protein (GFP) tagged p53 were incubated with and without DNA damaging agents, cisplatin or eptoposide, which are widely used as chemotherapeutic drugs. Then, the diffusion coefficient of GFP-p53 was determined by RICS and it was significantly reduced after the drug treatment while that of the one without drug treatment was not. It is suggested that the drugs induced the interaction of p53 with either other proteins or DNA. Together, our results demonstrated that RICS is able to detect the protein dynamics which may be associated with protein-protein or protein-DNA interactions in living cells and it may be useful for the drug screening.

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用光栅图像相关光谱(RICS)测定活HeLa细胞中蛋白53的扩散系数。

我们应用光栅图像相关光谱(RICS)技术表征了DNA损伤剂处理前后活细胞中p53蛋白的动态变化。表达绿色荧光蛋白(GFP)标记p53的HeLa细胞与DNA损伤剂、顺铂或表topo苷(广泛用作化疗药物)一起或不一起孵育。然后通过RICS测定GFP-p53的扩散系数，药物治疗后GFP-p53的扩散系数明显降低，而未经药物治疗的GFP-p53的扩散系数则没有明显降低。提示这些药物诱导p53与其他蛋白或DNA相互作用。总之，我们的研究结果表明，RICS能够检测活细胞中可能与蛋白质-蛋白质或蛋白质- dna相互作用相关的蛋白质动力学，并可能对药物筛选有用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Journal of Biomaterials and Nanobiotechnology

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