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A Modified ELISA Method to Evaluate the Interaction of Schistosoma Mansoni Proteins With Plasminogen

A Modified ELISA Method to Evaluate the Interaction of Schistosoma Mansoni Proteins With Plasminogen

Luis G V Fernandes, Rafaela S Fernandes, Ana L T O Nascimento, Luciana C C Leite

Methods Mol Biol. 2020;2151:185-195.

PMID: 32452005

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP62354432	D-Val-Leu-Lys 4-nitroanilide dihydrochloride		62354-43-2	Price

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