A Unique Class of Cyclases with a Kinase Fold Catalyzes Enethiol-Mediated Macrocyclization of Aminovinyl-Cysteine Motifs in Lanthipeptides

作者：Xiangqian Xie, Wen Guo, Yin‐Zheng Xia, Lijuan Liao, Mengxin Sun, Jingxue Wang, Jiangtao Gao, Hongwei Yao, Huan Wang · 发表于：ACS Central Science · 年份：2025 · DOI：10.1021/acscentsci.5c00569 · 被引用次数：3 · 研究领域：Microbial Natural Products and Biosynthesis、Biochemical and Structural Characterization、Glycosylation and Glycoproteins Research

High Resolution Image Download MS PowerPoint Slide 2-Aminovinyl-cysteine (AviCys) motifs represent a unique class of macrocyclic structures found in many ribosomally synthesized and post-translationally modified peptides (RiPPs). Despite their essential role in bioactivity, their biosynthetic machinery, particularly the cyclases catalyzing Avi(Me)Cys macrocyclization, has not been fully characterized. Herein, we report the discovery and biosynthetic elucidation of class V lanthipeptides rosin A1–A3, which feature a lanthionine (Lan) macrocycle and a C-terminal 2-aminovinyl-3-methyl-cysteine (AviMeCys) macrocycle. Rosins promote the migration of human foreskin fibroblast (HSF) cells, representing the first examples of lanthipeptides with cell migration-promoting activity. Comprehensive in vitro reconstitution revealed that the regio- and stereoselective AviMeCys macrocyclization is catalyzed by RosX, a newly identified cyclase with a kinase-like fold. Therefore, RosX-like cyclases, originally misannotated as kinase-like proteins, represent a unique class of cyclases that utilize the enethiol group for AviCys/AviMeCys macrocyclization. Furthermore, we demonstrate that Lan formation in rosins follows a substrate-controlled cyclization pathway with kinetic acceleration by the complex of kinase RosK and lyase RosY, which is distinct from the AviMeCys macrocyclization. This study resolves the long-standing ambiguity of enzymatic AviCys macrocyclization and provides a basis for bios...