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Endocytosis and intracellular localisation of type 1 ribosome-inactivating protein saporin-s6. (Articolo in rivista)

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Endocytosis and intracellular localisation of type 1 ribosome-inactivating protein saporin-s6. (Articolo in rivista) (literal)

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Bolognesi A, Polito L, Scicchitano V, Orrico C, Pasquinelli G, Musiani S, Santi S, Riccio M, Bortolotti M, Battelli MG. (2012)
Endocytosis and intracellular localisation of type 1 ribosome-inactivating protein saporin-s6.
in Journal of Biological Regulators & Homeostatic Agents (Testo stamp.)
(literal)

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Bolognesi A, Polito L, Scicchitano V, Orrico C, Pasquinelli G, Musiani S, Santi S, Riccio M, Bortolotti M, Battelli MG. (literal)

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Department of Experimental Pathology, Alma Mater Studiorum - University of Bologna, Bologna, Italy; Department of Radiological and Histopathological Sciences, Alma Mater Studiorum - University of Bologna, Bologna, Italy; IGM-CNR, Unit of Bologna, c/o IOR, Bologna, Italy; Department of Anatomy and Histology, University of Modena and Reggio Emilia, Modena, Italy (literal)

Titolo

Endocytosis and intracellular localisation of type 1 ribosome-inactivating protein saporin-s6. (literal)

Abstract

Saporin-S6 is a single-chain ribosome-inactivating protein (RIP) that has low toxicity in cells and animals. When the protein is bound to a carrier that facilitates cellular uptake, the protein becomes highly and selectively toxic to the cellular target of the carrier. Thus, saporin-S6 is one of the most widely used RIPs in the preparation of immunoconjugates for anti-cancer therapy. The endocytosis of saporin-S6 by the neoplastic HeLa cells and the subsequent intracellular trafficking were investigated by confocal microscopy that utilises indirect immunofluorescence analysis and transmission electron microscopy that utilises a direct assay with gold-conjugated saporin-S6 and an indirect immunoelectron microscopy assay. Our results indicate that saporin-S6 was taken up by cells mainly through receptor-independent endocytosis. Confocal microscopy analysis showed around 30% co-localisation of saporin-S6 with the endosomal compartment and less than 10% co-localisation with the Golgi apparatus. The pathway identified by the immunofluorescence assay and transmission electron microscopy displayed a progressive accumulation of saporin-S6 in perinuclear vesicular structures. The main findings of this work are the following: i) the nuclear localisation of saporin-S6 and ii) the presence of DNA gaps resulting from basic sites in HeLa nuclei after intoxication with saporin-S6. (literal)

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