Department of Biochemistry, ICMR - Regional Medical Research Centre, Port Blair 744103, Andaman and Nicobar Islands, India.
Department of Plant Sciences, School of Biological Sciences, Madurai Kamraj University, Madurai, Tamil Nadu 625021, India.
Res Microbiol. 2023 Nov-Dec;174(8):104107. doi: 10.1016/j.resmic.2023.104107. Epub 2023 Jul 28.
Leptospirosis, a global reemerging zoonosis caused by the spirochete Leptospira, has severe human and veterinary implications. Cell wall hydrolase (LIC_10271) with LytM (peptidase M23) and LysM domains are found to be associated with various pathogenic bacteria. These domains regulate effects on extracellular matrix and biofilm components, which promote cell wall remodeling and pathogen dissemination in the host. In this study, we present the cloning, expression, purification, and characterization of LIC_10271. To determine the localization of LIC_10271 within the inner membrane of Leptospira, Triton X-114 subcellular fractionation and immunoblot studies were performed. Furthermore, r-LIC_10271 binds with peptidoglycan, lipopolysaccharide, and laminin in a dose-dependent manner. Analysis of the signal peptide, M23, and LysM domains revealed conservation primarily within the P1 group of Leptospira, which encompasses the most pathogenic species. Moreover, the presence of native-LIC_10271 in the inner membrane and the distribution of M23 and LysM domains across pathogenic strains indicates their potential involvement in the interaction between the host and Leptospira.
钩端螺旋体病是一种由螺旋体钩端螺旋体引起的全球性再现性动物传染病,对人类和兽医都有严重的影响。细胞壁水解酶 (LIC_10271) 具有 LytM(肽酶 M23)和 LysM 结构域,与各种致病性细菌有关。这些结构域调节对细胞外基质和生物膜成分的影响,促进宿主中细胞壁的重塑和病原体的传播。在本研究中,我们介绍了 LIC_10271 的克隆、表达、纯化和表征。为了确定 LIC_10271 在钩端螺旋体内膜中的定位,进行了 Triton X-114 亚细胞分级分离和免疫印迹研究。此外,r-LIC_10271 以剂量依赖的方式与肽聚糖、脂多糖和层粘连蛋白结合。信号肽、M23 和 LysM 结构域的分析表明,它们主要在包含最具致病性的物种的钩端螺旋体 P1 组内保守。此外,天然 LIC_10271 存在于内膜中,并且 M23 和 LysM 结构域在致病性菌株中的分布表明它们可能参与宿主与钩端螺旋体之间的相互作用。