Cell surface nucleolin interacts with and internalizes Bothrops asper Lys49 phospholipase A2 and mediates its toxic activity
artículo original
Fecha
2018Autor
Lina Massimino, Maria
Simonato, Morena
Spolaore, Barbara
Franchin, Cinzia
Arrigoni, Giorgio
Marin, Oriano
Monturiol Gross, Laura
Fernández Ulate, Julián
Lomonte, Bruno
Tonello, Fiorella
Metadatos
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Phospholipases A2 are a major component of snake venoms. Some of them cause severe muscle necrosis
through an unknown mechanism. Phospholipid hydrolysis is a possible explanation of their toxic action,
but catalytic and toxic properties of PLA2s are not directly connected. In addition, viperid venoms
contain PLA2-like proteins, which are very toxic even if they lack catalytic activity due to a critical
mutation in position 49. In this work, the PLA2-like Bothrops asper myotoxin-II, conjugated with the
fluorophore TAMRA, was found to be internalized in mouse myotubes, and in RAW264.7 cells. Through
experiments of protein fishing and mass spectrometry analysis, using biotinylated Mt-II as bait, we
found fifteen proteins interacting with the toxin and among them nucleolin, a nucleolar protein present
also on cell surface. By means of confocal microscopy, Mt-II and nucleolin were shown to colocalise, at
4 °C, on cell membrane where they form Congo-red sensitive assemblies, while at 37 °C, 20 minutes after
the intoxication, they colocalise in intracellular spots going from plasmatic membrane to paranuclear
and nuclear area. Finally, nucleolin antagonists were found to inhibit the Mt-II internalization and toxic
activity and were used to identify the nucleolin regions involved in the interaction with the toxin
External link to the item
10.1038/s41598-018-28846-4Colecciones
- Microbiología [1171]
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