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A soluble form of Sda-β1,4-N-acetylgalactosaminyltransferase is released by differentiated human colon carcinoma CaCo-2 cells

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Abstract

We have previously shown that human colon carcinoma CaCo-2 cells express the Sda-β1,4-N-acetylgalactosaminyltransferase (Sda-βGalNAc-transferase) and that the enzyme activity correlates with the degree of enterocytic differentiation. Here we report that a large amount of this glycosyltransferase is released in soluble form, particularly when CaCo-2 cells are maintained in culture for more than 3 weeks in order to ensure an higher degree of enterocyte differentiation. The soluble enzyme was concentrated and partially purified by Blue-Sepharose and fetuin-Sepharose chromatography. The substrate specificity of the partially purified enzyme was similar to that of Sda-enzyme from epithelial cells of colon mucosa, and for its activity strictly required the presence in acceptors of NeuAc in α2,3-linkage to subterminal galactose. Among the low molecular glycans tested, NeuAcα2,3Galβ1,4GlcNAc appeared to be the best acceptor, whereas sialyl-Lewisx and sialyl-Lewisa did not serve as acceptors, indicating that the fucosylation of sub-terminal GlcNAc hindered the transferase activity. Contrary to this, the activity towards a disialylated acceptor such as di-sialyl-lacto-N-tetraose was reduced but not abolished. When CaCo-2 cells were cultured on porous membranes and the transferase activity assayed in medium collected from chambers corresponding to either the apical or basolateral face of highly differentiated CaCo-2 cells, a preferential release from the basolateral surface was found. Considering that Sda-βGalNAc-transferase is mainly located in the large intestine, current results support the notion that colonic cells largely contribute to the presence of the enzyme in human plasma.

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Authors and Affiliations

  1. Dipartimento di Patologia sperimentale, Università di Bologna, Via S. Giacomo 14, 40126, Bologna, Italy

    Franca Serafini-Cessi, Nadia Malagolini, Stefania Guerrini & Ileana Turrini

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  1. Franca Serafini-Cessi
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  2. Nadia Malagolini
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  3. Stefania Guerrini
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  4. Ileana Turrini
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Serafini-Cessi, F., Malagolini, N., Guerrini, S. et al. A soluble form of Sda-β1,4-N-acetylgalactosaminyltransferase is released by differentiated human colon carcinoma CaCo-2 cells. Glycoconjugate J 12, 773–779 (1995). https://doi.org/10.1007/BF00731238

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  • DOI: https://doi.org/10.1007/BF00731238

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