@article{3c92b84b036a4190aef27bb44e5fa16a,
title = "Structures of the holoenzyme TglHI required for 3-thiaglutamate biosynthesis",
abstract = "Structural diverse natural products like ribosomally synthesized and posttranslationally modified peptides (RiPPs) display a wide range of biological activities. Currently, the mechanism of an uncommon reaction step during the biosynthesis of 3-thiaglutamate (3-thiaGlu) is poorly understood. The removal of the β-carbon from the Cys in the TglA-Cys peptide catalyzed by the TglHI holoenzyme remains elusive. Here, we present three crystal structures of TglHI complexes with and without bound iron, which reveal that the catalytic pocket is formed by the interaction of TglH–TglI and that its activation is conformation dependent. Biochemical assays suggest a minimum of two iron ions in the active cluster, and we identify the position of a third iron site. Collectively, our study offers insights into the activation and catalysis mechanisms of the non-heme dioxygen-dependent holoenzyme TglHI. Additionally, it highlights the evolutionary and structural conservation in the DUF692 family of biosynthetic enzymes that produce diverse RiPPs.",
keywords = "RiPP, TglHI holoenzyme, X-ray, mechanism, metalloenzymes",
author = "Yanhui Zheng and Xiaoqing Xu and Xiaoli Fu and Xuerong Zhou and Chao Dou and Yue Yu and Weizhu Yan and Jingyuan Yang and Minqin Xiao and {van der Donk}, {Wilfred A.} and Xiaofeng Zhu and Wei Cheng",
note = "We thank the staff of the SSRF BL18U1 and BL02U1 for X-ray diffraction data collection, and we are grateful to Professor Yijun Gu for data collection and handling. We thank Dr. Guanglei Cheng, Dr. Jiahui Yang and Dr. Xi Wu (Analytical & Testing Center, SCU) for ICP-OES work. We thank the Core Facilities of West China Hospital for MS work and are grateful to Dan Du, Sisi Wu and Qianlun Pu for data analysis. This work was funded by the National Science Fund for Distinguished Young Scholars ( 32225001 ), the National Natural Science Foundation of China ( 81930125 ) and the 1.3.5 Project for Disciplines Excellence of West China Hospital, Sichuan University ( ZYYC20005 ), Sichuan Province Science and Technology Support Program ( 2021JDRC0029 ), National Natural Science Foundation of China ( 32200117 ), China Postdoctoral Science Foundation ( 2022M722281 ), Natural Science Foundation of Sichuan Province of China ( 2023NSFSC1197 ), the Fundamental Research Funds for the Central Universities ( 2022SCU12036 ) and the Howard Hughes Medical Institute . We thank the staff of the SSRF BL18U1 and BL02U1 for X-ray diffraction data collection, and we are grateful to Professor Yijun Gu for data collection and handling. We thank Dr. Guanglei Cheng, Dr. Jiahui Yang and Dr. Xi Wu (Analytical & Testing Center, SCU) for ICP-OES work. We thank the Core Facilities of West China Hospital for MS work and are grateful to Dan Du, Sisi Wu and Qianlun Pu for data analysis. This work was funded by the National Science Fund for Distinguished Young Scholars (32225001), the National Natural Science Foundation of China (81930125) and the 1.3.5 Project for Disciplines Excellence of West China Hospital, Sichuan University (ZYYC20005), Sichuan Province Science and Technology Support Program (2021JDRC0029), National Natural Science Foundation of China (32200117), China Postdoctoral Science Foundation (2022M722281), Natural Science Foundation of Sichuan Province of China (2023NSFSC1197), the Fundamental Research Funds for the Central Universities (2022SCU12036) and the Howard Hughes Medical Institute. Conceptualization, W.C. and X. Zhu; Methodology, Y.Z. X.X. X.F. and X. Zhou; Investigation, Y.Z. X.X. X.F. X. Zhou, and C.D.; Validation, Y.Z. Y.Y. W.Y. J.Y. and M.X.; Writing—Original Draft, Y.Z.; Writing—Review & Editing, W.C. W.A.v.d.D. X. Zhu, Y.Z. C.D. and Y.Y.; Supervision, W.C. W.A.v.d.D. and X. Zhu; Funding Acquisition, W.C. X. Zhu, and C.D. The authors declare no competing interests. We support inclusive, diverse, and equitable conduct of research.",
year = "2023",
month = oct,
day = "5",
doi = "10.1016/j.str.2023.08.004",
language = "English (US)",
volume = "31",
pages = "1220--1232.e5",
journal = "Structure",
issn = "0969-2126",
publisher = "Cell Press",
number = "10",
}