Cucurbit[8]uril-mediated protein homotetramerization
Cucurbit[8]uril-mediated protein homotetramerization
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DOI：
10.1039/c2sc20625k
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发表时间：
2012-01-01
期刊：
影响因子：
8.4
通讯作者：
Brunsveld, Luc
中科院分区：
文献类型：
--
作者：
Dang, Dung T.;Schill, Jurgen;Brunsveld, Luc
已经设计了一种超分子蛋白质四聚体方法，其使得能够控制离散蛋白质四聚体的形成。超分子元素葫芦[8]脲已被用作蛋白质四聚体的诱导剂，结合蛋白质之间的内在亲和力，其以二聚体形式预组织蛋白质。研究中的荧光蛋白（dYFP，dCFP）上的二聚化界面与遗传编码的N-末端苯丙氨酸-甘氨酸-甘氨酸（FGG）肽基序的组合允许葫芦[8]脲选择性诱导FGG-dYFP或FGG-dCFP四聚化。通过荧光各向异性、动态光散射和体积排阻色谱实验的结合，阐明了葫芦[8]脲诱导蛋白质四聚化的概念。葫芦[8]脲诱导的四聚体蛋白质复合物通过“二聚体的二聚体”途径形成，高度稳定，并且可以通过尺寸排阻色谱法分离。这种超分子诱导的蛋白质四聚体的方法开辟了一个新的入口，在产生定义明确的合成蛋白质组装。
A supramolecular protein tetramerization approach has been devised which enables the controlled formation of a discrete protein tetramer. The supramolecular element cucurbit[8]uril has been used as an inducer of the protein tetramerization in combination with intrinsic affinities between the proteins, which preorganize the protein in dimerized form. The combination of a dimerizing interface on the fluorescent proteins under study (dYFP, dCFP), with a genetically encoded N-terminal phenylalanine-glycine- glycine (FGG) peptide motif allows cucurbit[8]uril to selectively induce FGG-dYFP or FGG-dCFP tetramerization. The concept of cucurbit[8]uril-induced protein tetramerization was elucidated by using a combination of fluorescence anisotropy, dynamic light scattering and size exclusion chromatography experiments. The cucurbit[8]uril-induced tetrameric protein complex is formed via a "dimers of dimers'' pathway, is highly stable and can be separated by size exclusion chromatography. This supramolecular induced protein tetramerization approach opens up a novel entry in generating well-defined synthetic protein assemblies.