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Molecular Recognition of Surface Trans-Sialidases in Extracellular Vesicles of the Parasite Trypanosoma cruzi Using Atomic Force Microscopy (AFM).


ABSTRACT: Trans-sialidases (TS) are important constitutive macromolecules of the secretome present on the surface of Trypanosoma cruzi (T. cruzi) that play a central role as a virulence factor in Chagas disease. These enzymes have been related to infectivity, escape from immune surveillance and pathogenesis exhibited by this protozoan parasite. In this work, atomic force microscopy (AFM)-based single molecule-force spectroscopy is implemented as a suitable technique for the detection and location of functional TS on the surface of extracellular vesicles (EVs) released by tissue-culture cell-derived trypomastigotes (Ex-TcT). For that purpose, AFM cantilevers with functionalized tips bearing the anti-TS monoclonal antibody mAb 39 as a sense biomolecule are engineered using a covalent chemical ligation based on vinyl sulfonate click chemistry; a reliable, simple and efficient methodology for the molecular recognition of TS using the antibody-antigen interaction. Measurements of the breakdown forces between anti-TS mAb 39 antibodies and EVs performed to elucidate adhesion and forces involved in the recognition events demonstrate that EVs isolated from tissue-culture cell-derived trypomastigotes of T. cruzi are enriched in TS. Additionally, a mapping of the TS binding sites with submicrometer-scale resolution is provided. This work represents the first AFM-based molecular recognition study of Ex-TcT using an antibody-tethered AFM probe.

SUBMITTER: Prescilla-Ledezma A 

PROVIDER: S-EPMC9266976 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

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Molecular Recognition of Surface <i>Trans</i>-Sialidases in Extracellular Vesicles of the Parasite <i>Trypanosoma cruzi</i> Using Atomic Force Microscopy (AFM).

Prescilla-Ledezma Alexa A   Linares Fátima F   Ortega-Muñoz Mariano M   Retana Moreira Lissette L   Jódar-Reyes Ana Belén AB   Hernandez-Mateo Fernando F   Santoyo-Gonzalez Francisco F   Osuna Antonio A  

International journal of molecular sciences 20220628 13


<i>Trans</i>-sialidases (TS) are important constitutive macromolecules of the secretome present on the surface of <i>Trypanosoma cruzi</i> (<i>T. cruzi</i>) that play a central role as a virulence factor in Chagas disease. These enzymes have been related to infectivity, escape from immune surveillance and pathogenesis exhibited by this protozoan parasite. In this work, atomic force microscopy (AFM)-based single molecule-force spectroscopy is implemented as a suitable technique for the detection  ...[more]

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