School of Life Sciences, Tsinghua University, Beijing, China.
PLoS One. 2012;7(8):e44208. doi: 10.1371/journal.pone.0044208. Epub 2012 Aug 29.
In the present work, we developed the Fc-III tagged protein expression system for protein purification and detection. The Fc-III sequence encodes for a 13 residue peptide and this peptide is cyclized by disulfide bond formation when the fusion protein is expressed. The Fc-III-fusion proteins selectively bind to immunoglobulin Fc domains (IgG-Fc) expressed from E. coli. We showed the efficient purification of Fc-III tagged proteins by immobilized non-native IgG-Fc and the detection of the cellular locations of fusion proteins by fluorescent-conjugated IgG-Fc. Our results prove that Fc-III tagged protein expression system is a simple and efficient tool for protein purification and detection and is a useful addition to the biochemistry and proteomics toolbox.
在本工作中，我们开发了 Fc-III 标签蛋白表达系统，用于蛋白纯化和检测。Fc-III 序列编码一个 13 个氨基酸的肽段，当融合蛋白表达时，该肽段通过二硫键形成环化。Fc-III 融合蛋白特异性结合大肠杆菌表达的免疫球蛋白 Fc 结构域（IgG-Fc）。我们展示了通过固定化非天然 IgG-Fc 进行 Fc-III 标签蛋白的高效纯化，以及通过荧光标记 IgG-Fc 进行融合蛋白细胞定位的检测。我们的结果证明，Fc-III 标签蛋白表达系统是一种简单有效的蛋白纯化和检测工具，是生物化学和蛋白质组学工具包的有益补充。
