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利用双分子荧光互补测定法在酿酒酵母中进行蛋白质-蛋白质相互作用的体内定量。

In vivo quantification of protein-protein interactions in Saccharomyces cerevisiae using bimolecular fluorescence complementation assay.

机构信息

School of Biological Sciences and Research Center for Functional Cellulomics, Institute of Microbiology, Seoul National University, Seoul 151-747, Republic of Korea.

出版信息

J Microbiol Methods. 2010 Nov;83(2):194-201. doi: 10.1016/j.mimet.2010.08.021. Epub 2010 Sep 7.

Abstract

Most of the biological processes are carried out and regulated by dynamic networks of protein-protein interactions. In this study, we demonstrate the feasibility of the bimolecular fluorescence complementation (BiFC) assay for in vivo quantitative analysis of protein-protein interactions in Saccharomyces cerevisiae. We show that the BiFC assay can be used to quantify not only the amount but also the cell-to-cell variation of protein-protein interactions in S. cerevisiae. In addition, we show that protein sumoylation and condition-specific protein-protein interactions can be quantitatively analyzed by using the BiFC assay. Taken together, our results validate that the BiFC assay is a very effective method for quantitative analysis of protein-protein interactions in living yeast cells and has a great potential as a versatile tool for the study of protein function.

摘要

大多数生物过程都是通过蛋白质-蛋白质相互作用的动态网络来进行和调节的。在这项研究中,我们展示了双分子荧光互补(BiFC)测定法在活体定量分析酿酒酵母中蛋白质-蛋白质相互作用的可行性。我们表明,BiFC 测定法不仅可以定量分析蛋白质-蛋白质相互作用的量,还可以定量分析细胞间的变化。此外,我们还表明,通过使用 BiFC 测定法可以定量分析蛋白质的 sumoylation 和特定条件下的蛋白质-蛋白质相互作用。总之,我们的结果验证了 BiFC 测定法是一种非常有效的活体酵母细胞中蛋白质-蛋白质相互作用定量分析方法,并且作为研究蛋白质功能的通用工具具有很大的潜力。

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