Protein structure by mechanical triangulation
Protein structure by mechanical triangulation
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DOI：
10.1073/pnas.0509217103
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发表时间：
2006-01-31
影响因子：
11.1
通讯作者：
Rief, M
中科院分区：
文献类型：
--
作者：
Dietz, H;Rief, M
蛋白质结构的知识对于理解蛋白质功能至关重要。迄今为止，高分辨率蛋白质结构一直是集成方法的领域。在这里，我们开发了一种简单的单分子技术来测量溶液中折叠和功能蛋白质结构内选定残基的空间位置。具有受控连接拓扑结构的半胱氨酸工程化多蛋白的构建和机械展开允许以埃精度测量分子内距离。我们通过确定绿色荧光蛋白（GFP）结构中三个残基的位置来证明这种技术的潜力。我们的结果与GFP晶体结构完全一致。机械三角测量可以找到许多当前体结构方法失败的应用。
Knowledge of protein structure is essential to understand protein function. High-resolution protein structure has so far been the domain of ensemble methods. Here, we develop a simple single-molecule technique to measure spatial position of selected residues within a folded and functional protein structure in solution. Construction and mechanical unfolding of cysteine-engineered polyproteins with controlled linkage topology allows measuring intramolecular distance with angstrom precision. We demonstrate the potential of this technique by determining the position of three residues in the structure of green fluorescent protein (GFP). Our results perfectly agree with the GFP crystal structure. Mechanical triangulation can find many applications where current bulk structural methods fail.