Rational design of DNA sequence-specific zinc fingers.
Rational design of DNA sequence-specific zinc fingers.
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DNA序列特异性锌指的合理设计。
DOI：
10.1016/j.febslet.2012.02.025
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Sugiura Y.
中科院分区：
文献类型：
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作者：
Kono H;Imanishi M;Negi S;Tatsutani K;Sakaeda Y;Hashimoto A;Nakayama C;Futaki S;Sugiura Y.
我们开发了一个合理的计划，设计DNA结合蛋白。该方案被应用于锌指蛋白和设计的序列进行了实验，其特征在于具有高的DNA序列特异性。从已知指结构的骨架开始，我们最初计算出与预期结构相容的氨基酸序列，然后实验证实了所设计的指的二级结构。通过重新考虑氨基酸序列的一部分并计算选择具有最低蛋白质-DNA相互作用能的氨基酸，将DNA结合功能添加到所设计的指状物中。在设计的蛋白质中，一种蛋白质在最低和第二低蛋白质-DNA相互作用能之间具有足以提供DNA序列特异性的间隙。
We developed a rational scheme for designing DNA binding proteins. The scheme was applied for a zinc finger protein and the designed sequences were experimentally characterized with high DNA sequence specificity. Starting with the backbone of a known finger structure, we initially calculated amino acid sequences compatible with the expected structure and the secondary structures of the designed fingers were then experimentally confirmed. The DNA‐binding function was added to the designed finger by reconsidering a section of the amino acid sequence and computationally selecting amino acids to have the lowest protein–DNA interaction energy for the target DNA sequences. Among the designed proteins, one had a gap between the lowest and second lowest protein–DNA interaction energies that was sufficient to give DNA sequence‐specificity.