Custom tuning of Rieske oxygenase reactivity.
Custom tuning of Rieske oxygenase reactivity.
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DOI：
10.1038/s41467-023-41428-x
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发表时间：
2023-09-20
影响因子：
16.6
通讯作者：
Bridwell-Rabb, Jennifer
中科院分区：
文献类型：
--
作者：
Tian, Jiayi;Liu, Jianxin;Knapp, Madison;Donnan, Patrick H.;Boggs, David G.;Bridwell-Rabb, Jennifer
Rieske加氧酶使用Rieske型[2Fe-2S]簇和单核铁中心来启动一系列化学转化。然而，关于如何预测性地调整这种催化支架以催化不同的反应，存在很少的细节。因此，在这项工作中，使用结构分析的组合，以及基板和合理的蛋白质为基础的工程活动，我们阐明的建筑趋势，管理催化结果的Rieske单加氧酶TsaM。我们确定的结构特征，允许基板功能化的TsaM和精确的活性位点残基，可以有针对性地操纵反应。利用这些发现允许定制TsaM反应性的调节：鉴定支持不同TsaM催化反应的底物，并产生专门催化双氧化或顺序单氧化化学的变体。重要的是，我们进一步利用这些趋势来调整额外的单加氧酶和双加氧酶的反应性，从而提供定制调整Rieske加氧酶反应结果的策略。Rieske加氧酶化学对于生物化学途径是重要的，但如何预测性地调节常见的蛋白质支架以催化不同的反应仍然是难以捉摸的。在这里，作者报告了一种策略，可以合理地调整TsaM，一种Rieske单加氧酶，以催化双加氧和顺序单加氧反应，并定制其他Rieske加氧酶的反应性。
Rieske oxygenases use a Rieske-type [2Fe-2S] cluster and a mononuclear iron center to initiate a range of chemical transformations. However, few details exist regarding how this catalytic scaffold can be predictively tuned to catalyze divergent reactions. Therefore, in this work, using a combination of structural analyses, as well as substrate and rational protein-based engineering campaigns, we elucidate the architectural trends that govern catalytic outcome in the Rieske monooxygenase TsaM. We identify structural features that permit a substrate to be functionalized by TsaM and pinpoint active-site residues that can be targeted to manipulate reactivity. Exploiting these findings allowed for custom tuning of TsaM reactivity: substrates are identified that support divergent TsaM-catalyzed reactions and variants are created that exclusively catalyze dioxygenation or sequential monooxygenation chemistry. Importantly, we further leverage these trends to tune the reactivity of additional monooxygenase and dioxygenase enzymes, and thereby provide strategies to custom tune Rieske oxygenase reaction outcomes. Rieske oxygenase chemistry is important for biochemical pathways, but it remains elusive how a common protein scaffold can be predictively tuned to catalyze divergent reactions. Here, the authors report a strategy that can rationally tune TsaM, a Rieske monooxygenase to catalyze dioxygenation and sequential monooxygenation reactions, and customize the reactivity of other Rieske oxygenases.
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影响因子：
5.6
作者：
Dumitru, Razvan;Jiang, Wen Zhi;Weeks, Donald P.;Wilson, Mark A.
通讯作者：
Wilson, Mark A.
影响因子：
4.3
作者：
Csizi, Katja-Sophia;Eckert, Lina;Brunken, Christoph;Hofstetter, Thomas B.;Reiher, Markus
通讯作者：
Reiher, Markus
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
影响因子：
4.4
作者：
BECKE, AD
通讯作者：
BECKE, AD
DOI：
10.1021/acsenvironau.2c00023
发表时间：
2022-09-21
期刊：
ACS ENVIRONMENTAL AU
影响因子：
--
作者：
Bopp, Charlotte E;Bernet, Nora M;Kohler, Hans-Peter E;Hofstetter, Thomas B
通讯作者：
Hofstetter, Thomas B