Construction of a Blue Copper Analogue through Iterative Rational Protein Design Cycles Demonstrates Principles of Molecular Recognition in Metal Center Formation
Construction of a Blue Copper Analogue through Iterative Rational Protein Design Cycles Demonstrates Principles of Molecular Recognition in Metal Center Formation
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通过迭代理性蛋白质设计循环构建蓝铜类似物，展示了金属中心形成中的分子识别原理
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发表时间：
1998
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影响因子：
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通讯作者：
H. Hellinga
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作者：
H. Hellinga
蓝铜CysHis2Met主配位球位于硫氧还蛋白疏水核心内的不同方向的一系列蛋白质的构建和表征使人们能够探索蛋白质与金属之间的分子识别原理。一种自动化的Rational蛋白质设计算法预测了这些中心在结构上合适的位置，而不需要使用潜在的预先存在的结合位点，也不需要使用硫氧还蛋白宿主和已知的Blue Cu蛋白质之间的任何结构或序列同源性。构建了一系列的4个主要设计和32个变种。有必要用疏水壳层包围设计的初级配位球，以确保没有潜在的替代配位残基。要形成稳定的铜(II)-硫代键，需要破坏通常有利的氧化还原反应的不稳定性，在该反应中，硫醇被氧化成二硫化物。这是通过更深地埋藏配位的半胱氨酸来实现的。
The construction and characterization of a series of proteins in which the Blue Copper CysHis2Met primary coordination sphere was placed in various orientations within the hydrophobic core of thioredoxin has allowed exploration of the principles of molecular recognition between proteins and metals. An automated rational protein design algorithm predicted structurally suitable locations for these centers without use of either a potential preexisting binding site or any structural or sequence homology between the thioredoxin host and known Blue Copper proteins. A series of four primary designs and 32 variants were constructed. It was necessary to surround the designed primary coordination sphere with a hydrophobic shell to ensure the absence of potential alternative coordinating residues. Formation of a stable Cu(II)-thiolate bond required destabilization of a normally favored redox reaction in which the thiol is oxidized to a disulfide. This was achieved by more deeply burying the coordinating cysteine, pr...