Copper-dependent halogenase catalyses unactivated C-H bond functionalization

• 成果类型: Article
• 署名作者: Chiang, Chen-Yu; Ohashi, Masao; Le, Jessie; Chen, Pan-Pan; Zhou, Qingyang; Qu, Songrong; Bat-Erdene, Undramaa; Hematian, Shabnam; Rodriguez, Jose A.; Houk, K. N.; Guo, Yisong; Loo, Joseph A.; Tang, Yi
• 署名单位: University of California System; University of California Los Angeles; University of California System; University of California Los Angeles; University of North Carolina; University of North Carolina Greensboro; Carnegie Mellon University; University of California System; University of California Los Angeles
• 刊物名称: Nature
• ISSN/ISSBN: 0028-2101
• DOI: 10.1038/s41586-024-08362-4
• 发表日期: 2025-02-01
• 页码: 126-+
• 关键词: peptide ustiloxin b; biosynthesis; SPECTRA; hydroxylation; elucidation; proteins; atpenins; PATHWAY; FAMILY; genes

摘要

Carbon-hydrogen (C-H) bonds are the foundation of essentially every organic molecule, making them an ideal place to do chemical synthesis. The key challenge is achieving selectivity for one particular C(sp(3))-H bond(1, 2-3). In recent years, metalloenzymes have been found to perform C(sp(3))-H bond functionalization(4,5). Despite substantial progresses in the past two decades(6,7), enzymatic halogenation and pseudohalogenation of unactivated C(sp(3))-H-providing a functional handle for further modification-have been achieved with only non-haem iron/alpha-ketoglutarate-dependent halogenases, and are therefore limited by the chemistry possible with these enzymes(8). Here we report the discovery and characterization of a previously unknown halogenase ApnU, part of a protein family containing domain of unknown function 3328 (DUF3328). ApnU uses copper in its active site to catalyse iterative chlorinations on multiple unactivated C(sp(3))-H bonds. By taking advantage of the softer copper centre, we demonstrate that ApnU can catalyse unprecedented enzymatic C(sp(3))-H bond functionalization such as iodination and thiocyanation. Using biochemical characterization and proteomics analysis, we identified the functional oligomeric state of ApnU as a covalently linked homodimer, which contains three essential pairs-one interchain and two intrachain-of disulfide bonds. The metal-coordination active site in ApnU consists of binuclear type II copper centres, as revealed by electron paramagnetic resonance spectroscopy. This discovery expands the enzymatic capability of C(sp(3))-H halogenases and provides a foundational understanding of this family of binuclear copper-dependent oxidative enzymes.