MPL36, a major plasminogen (PLG) receptor in pathogenicLeptospira,has an essential role during infection
Authors: Zhu Weinan , Passalia Felipe J. , Hamond Camila , Abe Cecília M. , Ko Albert I. , Barbosa Angela S. , Wunder Elsio A. 
Published: March 2023

AbstractLeptospirosis, a zoonosis with worldwide distribution, is caused by pathogenic spirochetes belonging to the genusLeptospira. 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 pathogenicLeptospira. Our results showed that MPL36 is surface exposed and expressed during infection. Recombinant MPL36 (rMPL36) showed 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 ofmpl36has 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, ourmpl36mutant had a significant attenuated phenotype in the hamster model for acute leptospirosis. Our data indicates that MPL36 is the major PLG binding protein in pathogenicLeptospira, 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.Author SummaryAs part of their diverse virulence machinery, bacterial pathogens bind to human plasminogen (PLG) providing them with a proteolytic platform that promotes invasiveness, dissemination, and virulence. Leptospirosis is the leading zoonotic disease in morbidity and mortality worldwide.The burden of this neglected disease will continue to raise given the effects of climate change and social inequality, important drivers of disease. Furthermore, the gap of knowledge regarding leptospiral pathogenesis has negatively impacted the development of sensitive diagnostic tools and effective prevention methods. Previous studies have shown that pathogenicLeptospira, the causative agent of leptospirosis, can interact with PLG through different protein candidates. In this work, we characterized one of those candidates, Membrane Protein L36 (MPL36), as the main leptospiral plasminogen binding protein. Using genetically modified mutants,in vivo, andin vitroassays we provided evidence that MPL36 can bound PLG, promotes adherence to host cells and subsequent translocation, and degrades fibrinogen by converting bound-PLG to PLA, thus essential to leptospiral virulence. This work contributes to the growing field of bacterial pathogens exploring PLG to increase their virulence, while highlighting important new knowledge on leptospiral pathogenesis. MPL36 is an important candidate to be explored on the continued effort to improve diagnostic and prevention of this important zoonotic disease.