Human fibroblasts produce ceramide from sialyllactosylceramide on the plasma membranes. Sialidase Neu3 is known to be plasma membrane associated, while only indirect data suggest the plasma membrane association of β-galactosidase and β-glucosidase. To determine the presence of β-galactosidase and β-glucosidase on plasma membrane, cells were submitted to cell surface biotinylation. Biotinylated proteins were purified by affinity column and analyzed for enzymatic activities on artificial substrates. Both enzyme activities were found associated with the cell surface and were up-regulated in Neu3 overexpressing cells. These enzymes were capable to act on both artificial and natural substrates without any addition of activator proteins or detergents and displayed a trans activity in living cells.

Activity of plasma membrane β-galactosidase and β-glucosidase / M. Aureli, A.P. Masilamani, G. Illuzzi, N. Loberto, F. Scandroglio, A.E.G. Prinetti, V.L. Chigorno, S. Sonnino. - In: FEBS LETTERS. - ISSN 0014-5793. - 583:15(2009), pp. 2469-2473. [10.1016/j.febslet.2009.06.048]

Activity of plasma membrane β-galactosidase and β-glucosidase

M. Aureli
Primo
;
A.P. Masilamani
Secondo
;
G. Illuzzi;N. Loberto;F. Scandroglio;A.E.G. Prinetti;V.L. Chigorno
Penultimo
;
S. Sonnino
Ultimo
2009

Abstract

Human fibroblasts produce ceramide from sialyllactosylceramide on the plasma membranes. Sialidase Neu3 is known to be plasma membrane associated, while only indirect data suggest the plasma membrane association of β-galactosidase and β-glucosidase. To determine the presence of β-galactosidase and β-glucosidase on plasma membrane, cells were submitted to cell surface biotinylation. Biotinylated proteins were purified by affinity column and analyzed for enzymatic activities on artificial substrates. Both enzyme activities were found associated with the cell surface and were up-regulated in Neu3 overexpressing cells. These enzymes were capable to act on both artificial and natural substrates without any addition of activator proteins or detergents and displayed a trans activity in living cells.
β-Galactosidase; β-Glucosidase; Glycohydrolases; Plasma membrane; trans activity
Settore BIO/10 - Biochimica
Settore BIO/12 - Biochimica Clinica e Biologia Molecolare Clinica
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/71484
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