Tailorable Tetrahelical Bundles as a Toolkit for Redox Studies.
Tailorable Tetrahelical Bundles as a Toolkit for Redox Studies.
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DOI：
10.1021/acs.jpcb.2c05119
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发表时间：
2022-10-20
影响因子：
3.3
通讯作者：
Dutton, P. Leslie
中科院分区：
文献类型：
--
作者：
Solomon, Lee A.;Witten, Joshua;Kodali, Goutham;Moser, Christopher C.;Dutton, P. Leslie
氧化还原酶已经进化了数百万年，以执行对生命至关重要的各种代谢任务。理解这些任务是如何设计的依赖于提供外部电子供体或受体来启动电子转移反应。这是个挑战小分子氧化还原试剂可以不加选择地起作用，毒害细胞。天然氧化还原蛋白质更具选择性，但由于氧化还原电位的数量有限且难以调整，因此很难找到合适的伴侣。新生蛋白质提供了另一条途径，它们是强健的，可以承受突变，允许可定制的变化。它们也缺乏进化的人工产物，并且容易结合氧化还原辅因子。然而，还没有开发出一套可靠的工程原理，可以对这些蛋白质进行微调，使其氧化还原中点电位（Em）可以与任何天然氧化还原酶形成供体/受体对。这项工作解剖蛋白质辅因子的相互作用，可以调整到调节受体和供体的氧化还原电位，使用一个可变的从头设计的四螺旋蛋白质平台与铁四吡咯辅因子作为测试用例。我们展示了一系列的工程血红素b结合从头蛋白质和量化他们对EM的影响。通过关注表面电荷和掩埋电荷，以及辅因子的放置、化学修饰和辅因子的连接，我们能够实现跨越330 mV范围的宽范围Em值。我们预计这项工作将指导蛋白质工具的设计，这些工具可以与细胞内外的天然氧化还原酶相互作用，同时揭示天然蛋白质如何调节结合辅因子的Em值。
Oxidoreductases have evolved over millions of years to perform a variety of metabolic tasks crucial for life. Understanding how these tasks are engineered relies on delivering external electron donors or acceptors to initiate electron transfer reactions. This is a challenge. Small-molecule redox reagents can act indiscriminately, poisoning the cell. Natural redox proteins are more selective, but finding the right partner can be difficult due to the limited number of redox potentials and difficulty tuning them. De novo proteins offer an alternative path. They are robust and can withstand mutations that allow for tailorable changes. They are also devoid of evolutionary artifacts and readily bind redox cofactors. However, no reliable set of engineering principles have been developed that allow for these proteins to be fine-tuned so their redox midpoint potential (Em) can form donor/acceptor pairs with any natural oxidoreductase. This work dissects protein-cofactor interactions that can be tuned to modulate redox potentials of acceptors and donors using a mutable de novo designed tetrahelical protein platform with iron tetrapyrrole cofactors as a test case. We show a series of engineered heme b-binding de novo proteins and quantify their resulting effect on Em. By focusing on the surface charge and buried charges, as well as cofactor placement, chemical modification, and ligation of cofactors, we are able to achieve a broad range of Em values spanning a range of 330 mV. We anticipate this work will guide the design of proteinaceous tools that can interface with natural oxidoreductases inside and outside the cell while shedding light on how natural proteins modulate Em values of bound cofactors.
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影响因子：
16.6
作者：
Lichtenstein, Bruce R.;Bialas, Chris;Cerda, Jose F.;Fry, Bryan A.;Dutton, P. Leslie;Moser, Christopher C.
通讯作者：
Moser, Christopher C.
影响因子：
2.9
作者：
Calhoun, JR;Nastri, F;DeGrado, WF
通讯作者：
DeGrado, WF
影响因子：
2.9
作者：
Gibney, BR;Isogai, Y;Dutton, PL
通讯作者：
Dutton, PL
影响因子：
2.9
作者：
Gibney, BR;Rabanal, F;Dutton, PL
通讯作者：
Dutton, PL
影响因子：
14.8
作者：
Farid, Tammer A.;Kodali, Goutham;Solomon, Lee A.;Lichtenstein, Bruce R.;Sheehan, Molly M.;Fry, Bryan A.;Bialas, Chris;Ennist, Nathan M.;Siedlecki, Jessica A.;Zhao, Zhenyu;Stetz, Matthew A.;Valentine, Kathleen G.;Anderson, J. L. Ross;Wand, A. Joshua;Discher, Bohdana M.;Moser, Christopher C.;Dutton, P. Leslie
通讯作者：
Dutton, P. Leslie