Abstract
Leptospirosis is a widespread zoonotic disease caused by pathogenic Leptospira. Early and accurate diagnosis is the prime step in managing the disease. Secretory proteins of Leptospira remain distinguished for diagnosis due to their availability as soluble proteins in the serum and their interaction with the host immune response due to their extracellular presence. This study presents the cloning, expression, purification, and characterization of imelysin or LruB (LIC_10713), a putative leptospiral protein. We report that the localization of imelysin showed its presence in the inner membrane and in the culture supernatant. The imelysin was upregulated under in vitro physiological conditions of infection. The LIC_10713 interacted significantly with laminin, fibronectin, collagen type I, and collagen type IV in a dose-dependent manner. Phylogenetic analysis showed that LIC_10713 is predominately found in the pathogenic species of Leptospira, and the GxHxxE motif of imelysin-like proteins is represented as the amino acid sequence GWHAIE. Also, immunoglobulins in leptospirosis-infected patients recognize recombinant-LIC_10713 with 100% specificity and 90.9% sensitivity. The secretion nature, abundance, upregulation, binding to ECM components, and immunogenicity determine LIC_10713 as an important molecule that can be used as an anti-leptospirosis measure.
Key points
• The imelysin-like protein (LIC_10713) of Leptospira is a secretory protein
• The protein LIC_10713 can bind ECM molecules
• The LIC_10713 is mainly found in pathogenic leptospires
• The anti-LIC_10713 antibody from human serum can detect the r-LIC_10713
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The authors express their gratitude towards the Indian Council of Medical Research, New Delhi, India, for providing financial support under Grant No. Leptos/22/2013-ECD-I-2012–2400, and to the Department of Science and Technology, Science and Engineering Research Board, New Delhi, India, for providing financial assistance under Grant No. SR/SO/HS/0108/2012 to MGM. The authors also acknowledge the Indian Council of Medical Research, New Delhi, India, for awarding the Senior Research Fellowship through Grant No: Fellowship/131/2022-ECD-II-2021–8230 to AS and Grant No. ICMR-SRF 2020–0756/PROTEOMICS-BMS to HP.
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JT and MG conceived and designed the research; AS and DG conducted the experiments HP, AB, SH, AK, DB, and SN supported with reagents and resources, DB, SN JT, and MG supervision supervised and validated the research, AS wrote the draft, JT and MG reviewed and edited the manuscript. All authors read and approved the final manuscript.
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Sarma, A., Gunasekaran, D., Phukan, H. et al. Leptospiral imelysin (LIC_10713) is secretory, immunogenic and binds to laminin, fibronectin, and collagen IV. Appl Microbiol Biotechnol 107, 4275–4289 (2023). https://doi.org/10.1007/s00253-023-12573-6
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DOI: https://doi.org/10.1007/s00253-023-12573-6