De Novo Design of a Self-Assembled Artificial Copper Peptide that Activates and Reduces Peroxide.
De Novo Design of a Self-Assembled Artificial Copper Peptide that Activates and Reduces Peroxide.
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一种激活和减少过氧化物的自组装人工铜肽的从头设计。
DOI：
10.1021/acscatal.1c02132
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发表时间：
2021-08-20
期刊：
影响因子：
12.9
通讯作者：
Chakraborty, Saumen
中科院分区：
文献类型：
--
作者：
Mitra, Suchitra;Prakash, Divyansh;Rajabimoghadam, Khashayar;Wawrzak, Zdzislaw;Prasad, Pallavi;Wu, Tong;Misra, Sandeep K.;Sharp, Joshua S.;Garcia-Bosch, Isaac;Chakraborty, Saumen
含铜金属酶是一类重要的蛋白质，在自然界中催化许多反应。这类独特的金属酶的结构和功能特性的启发，我们报告的概念，设计，表征和功能研究的从头人工铜肽（ArCuP）内的三聚体自组装多肽支架，激活和减少过氧化物。使用第一原理方法，ArCuP被设计为经由在肽支架的a位点处引入的三个His残基来配位一个Cu。X-射线晶体学、紫外-可见光谱和电子顺磁共振（EPR）数据表明，Cu通过His的Nε原子结合形成T2 Cu环境。当与过氧化氢反应时，形成推定的铜-氢过氧物种，其中还原引发步骤加速其形成和还原的速率。质谱法被用来确定特定的残基进行氧化修饰，这表明他的氧化只在还原状态。用蛋白膜伏安法研究了ArCuP的氧化还原行为。的ArCuP的电催化行为的详细表征导致我们确定的催化参数（KM，kcat），建立了过氧化物酶活性的ArCuP。结合光谱和电化学数据表明pH依赖于反应性，这是最佳的pH 7.5。
Copper-containing metalloenzymes constitute a major class of proteins which catalyze a myriad of reactions in nature. Inspired by the structural and functional characteristics of this unique class of metalloenzymes, we report the conception, design, characterization, and functional studies of a de novo artificial copper peptide (ArCuP) within a trimeric self-assembled polypeptide scaffold that activates and reduces peroxide. Using a first principles approach, the ArCuP was designed to coordinate one Cu via three His residues introduced at an a site of the peptide scaffold. X-ray crystallographic, UV-vis and EPR data demonstrate that Cu binds via the Nε atoms of His forming a T2Cu environment. When reacted with hydrogen peroxide, the putative copper-hydroperoxo species is formed where a reductive priming step accelerates the rate of its formation and reduction. Mass spectrometry was used to identify specific residues undergoing oxidative modification, which showed His oxidation only in the reduced state. The redox behavior of the ArCuP was elucidated by protein film voltammetry. Detailed characterization of the electrocatalytic behavior of the ArCuP led us to determine the catalytic parameters (KM, kcat), which established the peroxidase activity of the ArCuP. Combined spectroscopic and electrochemical data showed a pH-dependence on the reactivity, which was optimum at pH 7.5.
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影响因子：
15
作者：
Hemsworth, Glyn R.;Taylor, Edward J.;Kim, Robbert Q.;Gregory, Rebecca C.;Lewis, Sally J.;Turkenburg, Johan P.;Parkin, Alison;Davies, Gideon J.;Walton, Paul H.
通讯作者：
Walton, Paul H.
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
DOI：
10.1073/pnas.1208822109
发表时间：
2012-11-13
影响因子：
11.1
作者：
Aachmann, Finn L.;Sorlie, Morten;Vaaje-Kolstad, Gustav
通讯作者：
Vaaje-Kolstad, Gustav
DOI：
10.1073/pnas.19.9.875
发表时间：
1933-01-01
影响因子：
11.1
作者：
Borsook, H;Keighley, G
通讯作者：
Keighley, G
影响因子：
14.8
作者：
Faiella, Marina;Andreozzi, Concetta;Lombardi, Angela
通讯作者：
Lombardi, Angela