Non-Native Anionic Ligand Binding and Reactivity in Engineered Variants of the Fe(II)- and α-Ketoglutarate-Dependent Oxygenase, SadA.
Non-Native Anionic Ligand Binding and Reactivity in Engineered Variants of the Fe(II)- and α-Ketoglutarate-Dependent Oxygenase, SadA.
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Fe（II）和α-酮戊二酸酯依赖性氧酶（Sada）的工程变体中的非母阴离子配体结合和反应性。
DOI：
10.1021/acs.inorgchem.2c02872
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发表时间：
2022-09-12
影响因子：
4.6
通讯作者：
Lewis, Jared C.
中科院分区：
文献类型：
--
作者：
Chan, Natalie H.;Gomez, Christian A.;Vennelakanti, Vyshnavi;Du, Qian;Kulik, Heather J.;Lewis, Jared C.
单核非血红素铁(II)-和α-酮戊二酸依赖加氧酶（FeDOs）催化位点选择性C-H羟基化。在某些情况下，这些酶的变体可以结合各种阴离子配体并催化非天然的氯化和溴化反应，其中Fe（II）结合面三联体中的保守的Asp/Glu残基被Ala/Gly取代。在这项研究中，我们探讨了不同阴离子与FeDO面部三元变体SadX的结合，以及这种结合对HO•与X•反弹的影响。我们发现氯化物和溴化物不仅能使非天然卤化反应发生，而且所研究的所有阴离子，包括叠氮化物、氰酸盐、甲酸盐和氟化物，都能显著加速和影响SadX羟基化催化的位点选择性。叠氮化物和氰酸盐也导致N3•、NCO•和OCN•反弹产物的形成。虽然没有观察到氟化物反弹，但该配体提供的速率加速使我们计算了从假定的Fe(III)(OH)(F)中间体反弹HO•和F•的势垒。这些计算表明，氟化的缺乏是由于HO•和F•反弹过渡态的相对障碍，而不是由于F•反弹无法进入的障碍。总之，这些结果提高了我们对FeDO面部三元变体对不同阴离子配体的耐受性的理解，它们促进涉及这些配体的反弹的能力，以及相对于HO•的固有反弹偏好，这将有助于利用这些酶开发非天然催化。在本研究中，我们探索了不同阴离子与Fe(II)-和α-酮戊二酸盐依赖的加氧酶的结合，其中Fe(II)-结合的Asp残基被Gly （SadX）取代，以及这种结合对HO•与X•反弹的影响。所研究的所有阴离子，包括氟化物，都能加速SadX羟基化催化，并观察到叠氮化物和氰酸盐反弹产生的产物。这些结果将有助于开发非原生的SadX催化剂。
Mononuclear non-heme Fe(II)- and α-ketoglutarate dependent oxygenases (FeDOs) catalyze site-selective C-H hydroxylation. Variants of these enzymes in which a conserved Asp/Glu residue in the Fe(II)-binding facial triad is replaced by Ala/Gly can, in some cases, bind various anionic ligands and catalyze non-native chlorination and bromination reactions. In this study, we explore the binding of different anions to a FeDO facial triad variant, SadX, and the effects of that binding on HO• vs. X• rebound. We establish that chloride and bromide not only enable non-native halogenation reactions but that all anions investigated, including azide, cyanate, formate, and fluoride, significantly accelerate and influence the site selectivity of SadX hydroxylation catalysis. Azide and cyanate also lead to the formation of products resulting from N3•, NCO•, and OCN• rebound. While fluoride rebound is not observed, the rate acceleration provided by this ligand led us to calculate barriers for HO• and F• rebound from a putative Fe(III)(OH)(F) intermediate. These calculations suggest that the lack of fluorination is due to the relative barriers of the HO• and F• rebound transition states rather than an inaccessible barrier for F• rebound. Together, these results improve our understanding of FeDO facial triad variant tolerance of different anionic ligands, their ability to promote rebound involving those ligands, and inherent rebound preferences relative to HO• that will aid efforts to develop non-native catalysis using these enzymes. In this study, we explore the binding of different anions to a Fe(II)- and α-ketoglutarate dependent oxygenase in which a Fe(II)-binding Asp residue is replaced by Gly (SadX), and the effects of that binding on HO• vs. X• rebound. All anions investigated, including fluoride, accelerate SadX hydroxylation catalysis, and products resulting from both azide and cyanate rebound are observed. These results will aid efforts to develop non-native SadX catalysis.
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影响因子：
3.7
作者：
Qin HM;Miyakawa T;Jia MZ;Nakamura A;Ohtsuka J;Xue YL;Kawashima T;Kasahara T;Hibi M;Ogawa J;Tanokura M
通讯作者：
Tanokura M
影响因子：
4.5
作者：
Papadopoulou, Athena;Meierhofer, Jasmin;Buller, Rebecca
通讯作者：
Buller, Rebecca
DOI：
10.1073/pnas.0909649106
发表时间：
2009-10-20
影响因子：
11.1
作者：
Matthews, Megan L.;Neumann, Christopher S.;Bollinger, J. Martin, Jr.
通讯作者：
Bollinger, J. Martin, Jr.
影响因子：
3.2
作者：
Straganz, G. D.;Nidetzky, B.
通讯作者：
Nidetzky, B.
影响因子：
14.8
作者：
Matthews, Megan L.;Chang, Wei-chen;Layne, Andrew P.;Miles, Linde A.;Krebs, Carsten;Bollinger, J. Martin, Jr.
通讯作者：
Bollinger, J. Martin, Jr.