异黄酮葡萄糖苷丰根霉素是一种针对 A 型分拣酶和溶解松脂素的抑制剂，能够减轻李斯特菌单胞菌在体内和体外的毒力。
Isoflavone glucoside genistin, an inhibitor targeting Sortase A and Listeriolysin O, attenuates the virulence of Listeria monocytogenes in vivo and in vitro.
发表日期：2023 Feb 04
作者：
Minda Liu, Qianghua Lv, Jingwen Xu, Baichen Liu, Yonglin Zhou, Siqi Zhang, Xue Shen, Lin Wang
来源：
BIOCHEMICAL PHARMACOLOGY
摘要：
作为一种常见的细胞内兼性革兰氏阳性细菌，单胞杆菌（Listeria monocytogenes, L. monocytogenes）展现出强大的抵抗能力，能够在极端环境如低温和广泛pH值范围内存活，从而在食品生产和加工中造成污染。而SrtA和LLO是L. monocytogenes的两个关键毒力因子，被广泛认为是抗L. monocytogenes感染药物开发的潜在靶点。在本研究中，我们发现异黄酮苷可以同时抑制SrtA的肽酶活性和LLO的溶血活性，而不影响L. monocytogenes的生长，从而减轻了开发抗药性的担忧。此外，我们证明异黄酮苷可以减少L. monocytogenes的生物膜形成和入侵人类结肠癌（Caco-2）细胞。随后的机制研究表明，异黄酮苷通过阻止LLO的寡聚化来抑制LLO介导的Caco-2细胞损伤。荧光淬灭法揭示了SrtA和LLO与异黄酮苷的潜在结合模式。异黄酮苷可能通过Leu33、Asn29和Met40残基结合到SrtA的活性口袋，并通过Asn259残基与LLO的D1域相互作用，参与寡聚化和孔形成过程。感染模型研究表明，异黄酮苷可以减少L. monocytogenes感染小鼠时的死亡率和病理损害。这些结果表明，异黄酮苷是一种有前途的抗毒力剂，可被考虑作为治疗L. monocytogenes感染的替代候选药物。版权所有©2023作者。由Elsevier Inc.出版。保留所有权利。
As a common intracellular facultative anaerobic Gram-positive bacterium, Listeria monocytogenes (L. monocytogenes) exhibits strong resistance to extreme environments, such as low temperature and a wide range of pH values, causing contamination in food production and processing. Sortase A (SrtA) and listeriolysin O (LLO), two crucial virulence factors of L. monocytogenes, are widely recognized as potential targets for the development of anti-L. monocytogenes infection drugs. In this study, we found that genistin simultaneously inhibits the peptidase activity of SrtA and the hemolytic activity of LLO without affecting the growth of L. monocytogenes, alleviating concerns about developing resistance. Furthermore, we demonstrated that genistin reduces L. monocytogenes biofilm formation and invasion of human colorectal cancer (Caco-2) cells. Subsequent mechanistic studies revealed that genistin inhibited LLO-mediated Caco-2 cell damage by blocking LLO oligomerization. Fluorescence quenching assay revealed the potential binding mode of SrtA and LLO to genistin. Genistin might bind to the active pocket of SrtA through residues Leu33, Asn29, and Met40, interacting with D1 domain of LLO involved in oligomerization and pore formation through residues Asn259. Studies in infection models revealed that genistin reduces mortality and pathological damage in mice infected with L. monocytogenes. These results indicate that genistin is a promising anti-virulence agent that could be considered an alternative candidate for the treatment of L. monocytogenes infection.Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.