文献类型：期刊文献

英文题名：Dissecting the Mechanism of the Nonheme Iron Endoperoxidase FtmOx1 Using Substrate Analogues

作者：Zhu, Guoliang[1];Yan, Wupeng[2];Wang, Xinye[1];Cheng, Ronghai[3];Naowarojna, Nathchar[3];Wang, Kun[1];Wang, Jun[2];Song, Heng[4];Wang, Yuyang[1];Liu, Hairong[5];Xia, Xuekui[5];Costello, Catherine E.[3];Liu, Xueting[1];Zhang, Lixin[1];Liu, Pinghua[3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200237, Peoples R China;[3]Boston Univ, Dept Chem, Boston, MA 02215 USA;[4]Wuhan Univ, Coll Chem & Mol Sci, Wuhan 430072, Hubei, Peoples R China;[5]Qilu Univ Technol, Inst Biol, Key Biosensor Lab Shandong Prov, Shandong Acad Sci, Jinan 250013, Shandong, Peoples R China

年份：2022

卷号：2

期号：7

起止页码：1686

外文期刊名：JACS AU

收录：EI(收录号：20225013244479);WOS:【ESCI(收录号:WOS:000819222300001)】；

基金：This work was supported by the National Natural Science Foundation of China (31720103901), the National Key Research and Development Program of China (2020YFA090032 and 2019YFA0906201), the National Natural Science Foundation of China (21977029 and 81903529), China Postdoctoral Science Foundation funded project (2019M661403), the Open Project Funding of the State Key Laboratory of Bioreactor Engineering, the 111 Project (B18022), and Shanghai Science and Technology Commission (18JC1411900). The authors thank staffs from BL02U1 and BL19U1 beamline at Shanghai Synchrotron Radiation Facility (SSRF), China, for their assistance in X-ray crystal data collection. This work is supported in part by the National Institutes of Health (GM140040 to P.L. and R24 GM134210 to C.E.C.).

语种：英文

外文关键词：FtmOx1; nonheme iron enzymes; endoperoxidation; hydroxylation; dehydrogenation; X-ray structure

摘要：FtmOx1 is a nonheme iron (NHFe) endoperoxidase, catalyzing three disparate reactions, endoperoxidation, alcohol dehydrogenation, and dealkylation, under in vitro conditions; the diversity complicates its mechanistic studies. In this study, we use two substrate analogues to simplify the FtmOx1-catalyzed reaction to either a dealkylation or an alcohol dehydrogenation reaction for structure-function relationship analysis to address two key FtmOx1 mechanistic questions: (1) Y224 flipping in the proposed COX-like model vs alpha-ketoglutarate (alpha KG) rotation proposed in the CarC-like mechanistic model and (2) the involvement of a Y224 radical (COX-like model) or a Y68 radical (CarC-like model) in FtmOx1-catalysis. When 13-oxo-fumitremorgin B (7) is used as the substrate, FtmOx1-catalysis changes from the endoperoxidation to a hydroxylation reaction and leads to dealkylation. In addition, consistent with the dealkylation side-reaction in the COX-like model prediction, the X-ray structure of the FtmOx1 center dot Co-II center dot alpha KG center dot 7 ternary complex reveals a flip of Y224 to an alternative conformation relative to the FtmOx1 center dot Fe-II center dot alpha KG binary complex. Verruculogen (2) was used as a second substrate analogue to study the alcohol dehydrogenation reaction to examine the involvement of the Y224 radical or Y68 radical in FtmOx1-catalysis, and again, the results from the verruculogen reaction are more consistent with the COX-like model.