Molecular Recognition of Surface Trans-Sialidases in Extracellular Vesicles of the Parasite Trypanosoma cruzi Using Atomic Force Microscopy (AFM)
Metadatos
Mostrar el registro completo del ítemAutor
Prescilla Ledezma, Alexa; Linares Ordóñez, Fátima; Ortega Muñoz, Mariano; Retana Moreira, Lissette; Jódar Reyes, Ana Belén; Hernández Mateo, Fernando; Santoyo González, Francisco; Osuna Carrillo De Albornoz, AntonioEditorial
MDPI
Fecha
2022-06-28Referencia bibliográfica
Prescilla-Ledezma, A... [et al.]. Molecular Recognition of Surface Trans-Sialidases in Extracellular Vesicles of the Parasite Trypanosoma cruzi Using Atomic Force Microscopy (AFM). Int. J. Mol. Sci. 2022, 23, 7193. [https://doi.org/10.3390/ijms23137193]
Patrocinador
ERANet program ERANet17/HLH-0142; Spanish Government PGC2018-099424-B-I00Resumen
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.