De Novo Design, Solution Characterization, and Crystallographic Structure of an Abiological Mn-Porphyrin-Binding Protein Capable of Stabilizing a Mn(V) Species.
De Novo Design, Solution Characterization, and Crystallographic Structure of an Abiological Mn-Porphyrin-Binding Protein Capable of Stabilizing a Mn(V) Species.
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能够稳定Mn（V）物种的非生物Mn-卟啉结合蛋白的从头设计、溶液表征和晶体结构。
DOI：
10.1021/jacs.0c10136
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发表时间：
2021-01-13
影响因子：
15
通讯作者：
DeGrado WF
中科院分区：
文献类型：
--
作者：
Mann SI;Nayak A;Gassner GT;Therien MJ;DeGrado WF
从头开始的蛋白质设计提供了测试我们对金属蛋白质如何执行困难转化的理解的机会。获得高分辨率的结构信息对于理解这种设计如何发挥作用至关重要。在设计卟啉结合蛋白方面已经取得了许多成功；然而，晶体特征一直难以捉摸，限制了从这类研究中获得的知识以及对新功能的扩展。此外，高氧化性高价中间体的形成带来了以前没有实现的设计挑战：(1)有目的地设计底物/氧化剂进入结合部位和(2)限制蛋白质支架的有害氧化。在这里，我们报道了第一个结晶学特征的卟啉结合蛋白，它被编程为不仅与合成的锰卟啉结合，而且保持结合部位的访问以形成高价氧化态。我们在Mn中心的开放配位位置显式地设计了一个可与氧单元结合的结合部位。在溶液中，蛋白质能够接触到高价的Mn(V)-oxo物种，该物种可以将O原子转移到硫醚底物上。晶体结构在设计的0.6ó以内，实际上包含一个水配体和第二个水分子，第二个水分子通过氢键稳定在活性中心的Gln侧链上，为观察到的反应活性提供了结构解释。
De novo protein design offers the opportunity to test our understanding of how metalloproteins perform difficult transformations. Attaining high-resolution structural information is critical to understanding how such designs function. There have been many successes in the design of porphyrin-binding proteins; however, crystallographic characterization has been elusive, limiting what can be learned from such studies as well as the extension to new functions. Moreover, formation of highly oxidizing high-valent intermediates poses design challenges that have not been previously implemented: (1) purposeful design of substrate/oxidant access to the binding site and (2) limiting deleterious oxidation of the protein scaffold. Here we report the first crystallographically characterized porphyrin-binding protein that was programmed to not only bind a synthetic Mn–porphyrin but also maintain binding site access to form high-valent oxidation states. We explicitly designed a binding site with accessibility to dioxygen units in the open coordination site of the Mn center. In solution, the protein is capable of accessing a high-valent Mn(V)–oxo species which can transfer an O atom to a thioether substrate. The crystallographic structure is within 0.6 Å of the design and indeed contained an aquo ligand with a second water molecule stabilized by hydrogen bonding to a Gln side chain in the active site, offering a structural explanation for the observed reactivity.
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影响因子：
15
作者：
Mandimutsira, BS;Ramdhanie, B;Goldberg, DP
通讯作者：
Goldberg, DP
DOI：
10.1107/s0907444902003359
发表时间：
2002-05-01
期刊：
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
影响因子：
--
作者：
Hovmöller, S;Zhou, T;Ohlson, T
通讯作者：
Ohlson, T
影响因子：
62.1
作者：
Baglia RA;Zaragoza JPT;Goldberg DP
通讯作者：
Goldberg DP
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
影响因子：
7.7
作者：
Brandenberg OF;Fasan R;Arnold FH
通讯作者：
Arnold FH