Department of Epidemiology of Microbial Diseases, Yale School of Public Health, New Haven, Connecticut, United States of America.
Laboratory of Vaccine Development, Instituto Butantan, São Paulo, Brazil.
PLoS Pathog. 2023 Jul 24;19(7):e1011313. doi: 10.1371/journal.ppat.1011313. eCollection 2023 Jul.
Leptospirosis, a zoonosis with worldwide distribution, is caused by pathogenic spirochetes belonging to the genus Leptospira. Bacterial outer membrane proteins (OMPs), particularly those with surface-exposed regions, play crucial roles in pathogen dissemination and virulence mechanisms. Here we characterized the leptospiral Membrane Protein L36 (MPL36), a rare lipoprotein A (RlpA) homolog with a C-terminal Sporulation related (SPOR) domain, as an important virulence factor in pathogenic Leptospira. Our results confirmed that MPL36 is surface exposed and expressed during infection. Using recombinant MPL36 (rMPL36) we also confirmed previous findings of its high plasminogen (PLG)-binding ability determined by lysine residues of the C-terminal region of the protein, with ability to convert bound-PLG to active plasmin. Using Koch's molecular postulates, we determined that a mutant of mpl36 has a reduced PLG-binding ability, leading to a decreased capacity to adhere and translocate MDCK cell monolayers. Using recombinant protein and mutant strains, we determined that the MPL36-bound plasmin (PLA) can degrade fibrinogen. Finally, our mpl36 mutant had a significant attenuated phenotype in the hamster model for acute leptospirosis. Our data indicates that MPL36 is the major PLG binding protein in pathogenic Leptospira, and crucial to the pathogen's ability to attach and interact with host tissues during infection. The MPL36 characterization contributes to the expanding field of bacterial pathogens that explore PLG for their virulence, advancing the goal to close the knowledge gap regarding leptospiral pathogenesis while offering a novel potential candidate to improve diagnostic and prevention of this important zoonotic neglected disease.
钩端螺旋体病是一种具有全球分布的人畜共患病,由属于钩端螺旋体属的致病性螺旋体引起。细菌外膜蛋白(OMPs),特别是那些具有暴露在表面的区域的 OMPs,在病原体传播和毒力机制中起着至关重要的作用。在这里,我们将钩端螺旋体膜蛋白 L36(MPL36)描述为一种重要的毒力因子,它是一种罕见的脂蛋白 A(RlpA)同源物,具有末端孢子相关(SPOR)结构域。我们的研究结果证实 MPL36 是表面暴露的,并在感染过程中表达。使用重组 MPL36(rMPL36),我们还证实了之前的发现,即该蛋白的 C 末端区域的赖氨酸残基决定了其对纤溶酶原(PLG)的高结合能力,并能够将结合的-PLG 转化为活性纤溶酶。使用 Koch 的分子假定,我们确定了 mpl36 的突变体具有降低的 PLG 结合能力,从而降低了粘附和易位 MDCK 细胞单层的能力。使用重组蛋白和突变株,我们确定 MPL36 结合的纤溶酶(PLA)可以降解纤维蛋白原。最后,我们的 mpl36 突变体在仓鼠急性钩端螺旋体病模型中表现出显著的减毒表型。我们的数据表明,MPL36 是致病性钩端螺旋体中主要的 PLG 结合蛋白,对于病原体在感染过程中附着和与宿主组织相互作用的能力至关重要。MPL36 的特性为探索 PLG 作为细菌病原体毒力的扩展领域做出了贡献,推进了缩小关于钩端螺旋体发病机制的知识差距的目标,同时提供了一种改进这种重要的人畜共患被忽视疾病的诊断和预防的新的潜在候选物。