Regional Medical Research Centre (ICMR), Post Bag No-13, Port Blair 744101, Andaman and Nicobar Islands, India; Department of Chemical Sciences, Ariel University, Ariel 70400, Israel.
Regional Medical Research Centre (ICMR), Post Bag No-13, Port Blair 744101, Andaman and Nicobar Islands, India.
Biochim Biophys Acta Proteins Proteom. 2018 May-Jun;1866(5-6):712-721. doi: 10.1016/j.bbapap.2018.04.006. Epub 2018 Apr 12.
Leptospira, the causative agent of leptospirosis is known to have many proteases with potential to degrade extracellular matrix. However, a multipronged approach to identify, classify, characterize and elucidate their role has not been attempted. Our proteomic approach using high-resolution LC-MS/MS analysis of Triton X-114 fractions of Leptospira interrogans resulted in the identification of 104 proteases out of 130 proteases predicted by MEROPS. In Leptospira approximately 3.5% of the genome complements for proteases, which include catalytic types of metallo-, serine-, cysteine-, aspartic-, threonine- and asparagine- peptidases. Comparison of proteases from different serovars revealed that M04, M09B, M14A, M75, M28A, A01 and U73 protease families are exclusively present in pathogenic form. The M23 and S33 protease families are represented with >14 members in Leptospira. The differential expression under physiological temperature (37 °C) and osmolarity (300 mOsM) showed that proteases belonging to the catalytic type of Metallo-peptidases are upregulated significantly in pathogenic conditions. In silico prediction and characterization of the proteases revealed that several proteases are membrane anchored and secretory, classical as well as non-classical system. The study demonstrates the diversity and complexity of proteases, while maintaining conservation across the serovars in Leptospira and their differential expression under pathogenic conditions.
钩端螺旋体是钩端螺旋体病的病原体，已知其具有许多潜在的能够降解细胞外基质的蛋白酶。然而，尚未尝试采用多管齐下的方法来鉴定、分类、表征和阐明其作用。我们使用 Triton X-114 fractions of Leptospira interrogans 的高分辨率 LC-MS/MS 分析的蛋白质组学方法，从 MEROPS 预测的 130 种蛋白酶中鉴定出 104 种蛋白酶。在钩端螺旋体中，大约 3.5%的基因组与蛋白酶互补，其中包括金属、丝氨酸、半胱氨酸、天冬氨酸、苏氨酸和天冬酰胺肽酶的催化类型。不同血清型的蛋白酶比较表明，M04、M09B、M14A、M75、M28A、A01 和 U73 蛋白酶家族仅存在于致病性形式中。M23 和 S33 蛋白酶家族在钩端螺旋体中有超过 14 个成员。生理温度（37°C）和渗透压（300 mOsM）下的差异表达表明，属于金属肽酶催化类型的蛋白酶在致病性条件下显著上调。蛋白酶的计算机预测和特征分析表明，几种蛋白酶是膜锚定和分泌的，包括经典和非经典系统。该研究表明了钩端螺旋体中蛋白酶的多样性和复杂性，同时在不同血清型之间保持了保守性，并在致病性条件下表现出差异表达。
