An expandable, modular de novo protein platform for precision redox engineering.
An expandable, modular de novo protein platform for precision redox engineering.
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DOI：
10.1073/pnas.2306046120
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发表时间：
2023-08
影响因子：
11.1
通讯作者：
Anderson, J. L. Ross
中科院分区：
文献类型：
--
作者：
Hutchins, George H.;Noble, Claire E. M.;Bunzel, H. Adrian;Williams, Christopher;Dubiel, Paulina;Yadav, Sathish K. N.;Molinaro, Paul M.;Barringer, Rob;Blackburn, Hector;Hardy, Benjamin J.;Parnell, Alice E.;Landau, Charles;Race, Paul R.;Oliver, Thomas A. A.;Koder, Ronald L.;Crump, Matthew P.;Schaffitzel, Christiane;Oliveira, A. Sofia F.;Mulholland, Adrian J.;Anderson, J. L. Ross
基于蛋白质的电路中的电子流动对生命至关重要，支持细胞能量产生和光合作用。虽然我们对这种天然电子传导机制的理解受益于结构基因组学时代的进步，但我们还没有完全利用这些生物电子组件和组件的特殊功能。为了直接解决这个问题，我们报告了一个可扩展的，模块化的蛋白质平台的设计，用于创建良好折叠的，新的自然蛋白质含有一个或多个氧化还原活性血红素辅因子。我们还表明，一个相对简单的计算设计策略可以用来扩展血红素含模块到一个7纳米的分子线，以及如何基本的生物物理性质的血红素在这样的蛋白质可以预测和操纵使用计算。生命的电子传导电路代表了一种尚未开发的精致的纳米级生物分子工程资源。在这里，我们报告的从头二血红素“模型”蛋白，4D2，我们随后使用创建一个扩展的，模块化的平台血红素蛋白设计的表征和结构。通过计算重新设计创建了一个折叠良好的单血红素变体，然后将其用于连续静电氧化还原电位计算的实验验证。这证明了如何预测和微调基本的生物物理特性。然后将4D2延伸成四血红素螺旋束，代表7 nm的分子线。尽管分子量仅为24 kDa，电子冷冻显微镜显示了显着的细节水平，表明二级结构和血红素辅因子的定位。这种强大的，可表达的，高度热稳定的和易于设计的模块化平台为氧化还原蛋白质设计和人工电子传导电路的未来构建提供了宝贵的资源。
The flow of electrons within protein-based circuitry is essential to life, underpinning cellular energy generation and photosynthesis. While our understanding of this natural electron-conducting machinery has benefitted from the advances of the structural genomics era, we have yet to fully exploit the exceptional features of these bioelectronic components and assemblies on our own terms. To directly address this, we report the design of an expandable, modular protein platform for creating well-folded, new-to-nature proteins containing one or more redox-active heme cofactors. We also demonstrate that a relatively simple computational design strategy can be used to extend heme-containing modules into a 7-nm molecular wire and how fundamental biophysical properties of hemes within such proteins can be predicted and manipulated using computation. The electron-conducting circuitry of life represents an as-yet untapped resource of exquisite, nanoscale biomolecular engineering. Here, we report the characterization and structure of a de novo diheme “maquette” protein, 4D2, which we subsequently use to create an expanded, modular platform for heme protein design. A well-folded monoheme variant was created by computational redesign, which was then utilized for the experimental validation of continuum electrostatic redox potential calculations. This demonstrates how fundamental biophysical properties can be predicted and fine-tuned. 4D2 was then extended into a tetraheme helical bundle, representing a 7 nm molecular wire. Despite a molecular weight of only 24 kDa, electron cryomicroscopy illustrated a remarkable level of detail, indicating the positioning of the secondary structure and the heme cofactors. This robust, expressible, highly thermostable and readily designable modular platform presents a valuable resource for redox protein design and the future construction of artificial electron-conducting circuitry.
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影响因子：
5.9
作者：
D'Souza, Areetha;Torres, Jaume;Bhattacharjya, Surajit
通讯作者：
Bhattacharjya, Surajit
影响因子：
8.4
作者：
Anderson JLR;Armstrong CT;Kodali G;Lichtenstein BR;Watkins DW;Mancini JA;Boyle AL;Farid TA;Crump MP;Moser CC;Dutton PL
通讯作者：
Dutton PL
影响因子：
15
作者：
Ghirlanda, G;Osyczka, A;DeGrado, WF
通讯作者：
DeGrado, WF
影响因子：
2.9
作者：
BASHFORD, D;KARPLUS, M
通讯作者：
KARPLUS, M
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K