A versatile platform for nanotechnology based on circular permutation of a chaperonin protein
A versatile platform for nanotechnology based on circular permutation of a chaperonin protein
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DOI：
10.1088/0957-4484/17/5/001
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发表时间：
2006-03-14
期刊：
影响因子：
3.5
通讯作者：
Trent, JD
中科院分区：
文献类型：
--
作者：
Paavola, CD;Chan, SL;Trent, JD
许多蛋白质复合物已被开发为纳米级模板。这些可以使用结合无机材料的肽序列或通过与结合蛋白或催化蛋白融合来功能化。为了将肽和蛋白质整合到蛋白质模板中的特定位置，我们使用循环排列来重新定位多肽链的氨基和羧基末端。然后可以将额外的序列连接到蛋白质末端。这最大限度地减少了对蛋白质结构的破坏，并减少了对添加序列的大小和构象的限制。我们将柴氏硫化叶菌伴侣蛋白亚基的末端重新定位到伴侣蛋白复合物外表面的五个不同位置（残基 153、267、316、480 和 499 之后）。这些变化将末端置于伴侣蛋白复合物的外表面上。这些排列形成双环和高阶组装，类似于在天然蛋白质中观察到的那些。当增强型黄色荧光蛋白与267和480位排列的两个伴侣蛋白亚基融合时，所得融合蛋白发出荧光并形成组装双环和高阶结构。该方法适用于其他纳米级蛋白质模板。
A number of protein complexes have been developed as nanoscale templates. These can be functionalized using peptide sequences that bind inorganic materials or by fusion to binding or catalytic proteins. In order to integrate peptides and proteins into specific positions in a protein template, we used circular permutation to relocate the amino and carboxy termini of the polypeptide chain. Additional sequences can then be joined to the protein termini. This minimizes disruption of the protein structure and reduces restrictions on size and conformation of the added sequence. We relocated the termini of a Sulfolobus shibatae chaperonin subunit to five different locations across the outside surface of the chaperonin complex (after residues 153, 267, 316, 480 and 499). These changes place the termini on the outside surface of the chaperonin complex. The permutants formed double rings and higher-order assemblies similar to those observed in the natural protein. When enhanced yellow fluorescent protein was fused to two chaperonin subunits permuted at positions 267 and 480, the resulting fusion protein was fluorescent and formed assembled double rings and higher-order structure. This approach is applicable to other nanoscale protein templates.