Gram-positive bacteria endow their peptidoglycan with glycopolymers that are crucial for viability and pathogenesis. However, the cellular machinery that executes this function is not well understood. While decades of genetic and phenotypic work have highlighted the LytR-CpsA-Psr (LCP) family of enzymes as cell-wall glycopolymer transferases, theirin vitrocharacterization has been elusive, largely due to a paucity of tools for functional assays. In this report, we synthesized authentic undecaprenyl diphosphate-linked wall teichoic acid (WTA) intermediates and built an assay system capable of monitoring LCP-mediated glycopolymer transfer. We report that allBacillus subtilisLCP enzymes anchor WTAs to peptidoglycanin vitro. Furthermore, we probed the catalytic requirements and substrate preferences for these LCP enzymes and elaboratedin vitroconditions for facile tests of enzyme function. This work sheds light on the molecular features of glycopolymer transfer and aims to aid drug discovery and development programs exploiting this promising antibacterial target.

中文翻译：

---

枯草芽孢杆菌LytR-CpsA-Psr酶从真实的脂质连接的底物壁壁酸性转移到成熟的肽聚糖体外。

革兰氏阳性细菌赋予其肽聚糖以对于生存能力和发病机理至关重要的糖聚合物。然而，执行该功能的蜂窝机械尚未被很好地理解。尽管数十年的遗传和表型研究突显了LytR-CpsA-Psr（LCP）酶家族作为细胞壁糖聚合物转移酶，但其体外表征一直难以捉摸，这主要是由于缺乏用于功能测定的工具。在本报告中，我们合成了正癸二烯基二磷酸十一碳二烯键联的壁壁chochochoic酸（WTA）中间体，并建立了能够监测LCP介导的糖聚合物转移的测定系统。我们报告所有枯草芽孢杆菌LCP酶锚定WTAs肽聚糖体外。此外，我们探讨了这些LCP酶的催化要求和底物偏好，并详细阐述了体外条件下酶功能的简便测试。这项工作揭示了糖聚合物转移的分子特征，旨在帮助开发利用这一有前途的抗菌目标的药物发现和开发计划。