一种重布线的绿色荧光蛋白：在非顺序、非环形 GFP 变体中的折叠和功能。
A rewired green fluorescent protein: folding and function in a nonsequential, noncircular GFP permutant.
机构信息
Department of Chemical and Biological Engineering, University of Colorado, Boulder, Colorado 80309, United States.
出版信息
Biochemistry. 2010 Dec 28;49(51):10773-9. doi: 10.1021/bi100975z. Epub 2010 Dec 3.
Abstract
The sequential order of secondary structural elements in proteins affects the folding and activity to an unknown extent. To test the dependence on sequential connectivity, we reconnected secondary structural elements by their solvent-exposed ends, permuting their sequential order, called "rewiring". This new protein design strategy changes the topology of the backbone without changing the core side chain packing arrangement. While circular and noncircular permutations have been observed in protein structures that are not related by sequence homology, to date no one has attempted to rationally design and construct a protein with a sequence that is noncircularly permuted while conserving three-dimensional structure. Herein, we show that green fluorescent protein can be rewired, still functionally fold, and exhibit wild-type fluorescence excitation and emission spectra.
摘要
蛋白质中二级结构元件的顺序对折叠和活性的影响程度未知。为了测试对顺序连接性的依赖性，我们通过溶剂暴露的末端重新连接二级结构元件，改变它们的顺序，称为“重布线”。这种新的蛋白质设计策略改变了骨架的拓扑结构，而不改变核心侧链堆积排列。虽然在序列同源性没有关系的蛋白质结构中已经观察到了圆形和非圆形的排列，但迄今为止，还没有人试图合理地设计和构建一种序列为非圆形排列但三维结构保守的蛋白质。在这里，我们表明绿色荧光蛋白可以被重新布线，仍然能够功能性折叠，并表现出野生型荧光激发和发射光谱。
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