Abstract
An important aspect of host-pathogen interactions is the interference of secreted proteins with the fibrinolytic system. Herein, we describe a modified ELISA method used to evaluate the interaction of a recombinant Schistosoma mansoni protein with plasminogen (PLG). Using this protocol, we demonstrated that a secreted protein, recombinant venom allergen-like protein 18 (rSmVAL18) acts as a plasminogen receptor increasing its activation into plasmin in the presence of the urokinase-type plasminogen activator (uPA). PLG binding was determined by immobilizing human PLG in the plate and incubating with the recombinant protein; competitive binding with a lysine analog demonstrated the interaction of the protein lysine residues with PLG Kringle domains. To assess the activation of S. mansoni recombinant protein-bound PLG, the amidolytic activity of generated plasmin was measured using the d-Val-Leu-Lys 4-nitroanilide dihydrochloride substrate.
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Acknowledgments
This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, grants 2012/23124-4, 2010/18486-9, 2014/50981-0 and 2017/06731-8), CNPq (grants 301229/2017-1 and 441449/2014-0) and Fundação Butantan.
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Fernandes, L.G.V., Fernandes, R.S., Nascimento, A.L.T.O., Leite, L.C.C. (2020). A Modified ELISA Method to Evaluate the Interaction of Schistosoma mansoni Proteins with Plasminogen. In: Timson, D.J. (eds) Schistosoma mansoni. Methods in Molecular Biology, vol 2151. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0635-3_15
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DOI: https://doi.org/10.1007/978-1-0716-0635-3_15
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