G(M2) Activator is a low molecular weight protein cofactor that stimulates the enzymatic conversion of G(M2) into G(M3) by human beta-hexosaminidase A and also the conversion of G(M2) into G(A2) by clostridial sialidase (Wu, Y.-Y., Lockyer, J. M., Sugiyama, E., Pavlova, N. V., Li, Y.-T., and Li, S.- C. (1994) J. Biol. Chem. 269, 16276-16283). Among the five known activator proteins for the enzymatic hydrolysis of glycosphingolipids, only G(M2) activator is effective in stimulating the hydrolysis of G(M2). However, the mechanism of action of G(M2) activator is still not well understood, Using a unique disialosylganglioside, GalNAc-G(D1a), as the substrate, we were able to show that in the presence of G(M2) activator, GalNAc-G(D1a) was specifically converted into GalNAc-G(M1a) by clostridial sialidase, while in the presence of saposin B, a nonspecific activator protein, GalNAc-G(D1a) was converted into both GalNAc-G(M1a) and GalNAc-G(M1b). individual products generated from GalNAc-G(D1a) by clostridial sialidase were identified by thin layer chromatography, negative secondary ion mass spectrometry, and immunostaining with a monoclonal IgM that recognizes the G(M2) epitope. Our results clearly show that G(M2) activator recognizes the G(M2) epitope in GalNAc-G(D1a). Thus, G(M2) activator may interact with the trisaccharide structure of the G(M2) epitope and render the GalNAc and NeuAc residues accessible to beta-hexosaminidase A and sialidase, respectively.

SPECIFIC RECOGNITION OF N-ACETYLNEURAMINIC ACID IN THE G(M2) EPITOPE BY HUMAN G(M2) ACTIVATOR PROTEIN / S. LI, Y. WU, E. SUGIYAMA, T. TAKI, T. KASAMA, R. CASELLATO, S. SONNINO, Y. LI. - In: THE JOURNAL OF BIOLOGICAL CHEMISTRY. - ISSN 0021-9258. - 270:41(1995), pp. 24246-24251.

SPECIFIC RECOGNITION OF N-ACETYLNEURAMINIC ACID IN THE G(M2) EPITOPE BY HUMAN G(M2) ACTIVATOR PROTEIN

R. CASELLATO;S. SONNINO
Penultimo
;
1995

Abstract

G(M2) Activator is a low molecular weight protein cofactor that stimulates the enzymatic conversion of G(M2) into G(M3) by human beta-hexosaminidase A and also the conversion of G(M2) into G(A2) by clostridial sialidase (Wu, Y.-Y., Lockyer, J. M., Sugiyama, E., Pavlova, N. V., Li, Y.-T., and Li, S.- C. (1994) J. Biol. Chem. 269, 16276-16283). Among the five known activator proteins for the enzymatic hydrolysis of glycosphingolipids, only G(M2) activator is effective in stimulating the hydrolysis of G(M2). However, the mechanism of action of G(M2) activator is still not well understood, Using a unique disialosylganglioside, GalNAc-G(D1a), as the substrate, we were able to show that in the presence of G(M2) activator, GalNAc-G(D1a) was specifically converted into GalNAc-G(M1a) by clostridial sialidase, while in the presence of saposin B, a nonspecific activator protein, GalNAc-G(D1a) was converted into both GalNAc-G(M1a) and GalNAc-G(M1b). individual products generated from GalNAc-G(D1a) by clostridial sialidase were identified by thin layer chromatography, negative secondary ion mass spectrometry, and immunostaining with a monoclonal IgM that recognizes the G(M2) epitope. Our results clearly show that G(M2) activator recognizes the G(M2) epitope in GalNAc-G(D1a). Thus, G(M2) activator may interact with the trisaccharide structure of the G(M2) epitope and render the GalNAc and NeuAc residues accessible to beta-hexosaminidase A and sialidase, respectively.
Settore BIO/10 - Biochimica
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/188754
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