G(M2) activator protein is a protein cofactor which stimulates the enzymatic hydrolysis of both GalNAc and NeuAc from G(M2). We have previously isolated two cDNA clones, G(M2) activator cDNA and G(M2A) cDNA, for human G(M2) activator protein (Nagarajan, S., Chen, H.-C., Li, S.-C., Li, Y.-T., and Lockyer, J. M. (1992) Biochem. J. 282, 807-813). G(M2A) mRNA is an RNA alternative splicing product that contains exons 1, 2, 3, and intron 3 of the genomic DNA sequence of G(M2) activator protein (Klima, H., Tanaka, A., Schnabel, D., Nakano, T., Schroder, M., Suzuki, K., and Sandhoff, K. (1991) FEES Left. 289, 260-264). G(M2A) cDNA encodes a protein (G(M2A) protein) containing 1-109 of the 160 amino acids of human G(M2) activator protein, plus a tripeptide (VST) encoded by intron 3 at the COOH terminus. Thus, G(M2A) protein can be regarded as a form (truncated version) of G(M2) activator protein. We have expressed G(M2A) cDNA in Escherichia coli using pT7-7 as the vector. The recombinant G(M2A) protein was purified to an electrophoretically homogeneous form and was found to stimulate the hydrolysis of NeuAc from G(M2) by clostridial sialidase, but not the hydrolysis of GalNAc from G(M2) by beta-hexosaminidase A. Like G(M2) activator protein, G(M2A) protein also specifically recognized the terminal G(M2) epitope in GalNAc-GD1a and stimulated the hydrolysis of only the external NeuAc from this ganglioside by clostridial sialidase. These results enabled us to discern the enzymatic hydrolyses of GalNAc and NeuAc from the G(M2) epitope and established that the NeuAc recognition domain of G(M2) activator protein is located within amino acids 1-109. The presence of G(M2A) mRNA in human tissues and the selective stimulation of NeuAc hydrolysis by G(M2A) protein indicate that this activator protein may be involved in the catabolism of G(M2) through the asialo-G(M2) pathway.

Characterization of an alternatively spliced G(M2) activator protein, G(M2A) protein - An activator protein which stimulates the enzymatic hydrolysis of N-acetylneuraminic acid, but not N-acetylgalactosamine, from G(M2) / Y. Wu, S. Sonnino, Y. Li, S. Li. - In: THE JOURNAL OF BIOLOGICAL CHEMISTRY. - ISSN 0021-9258. - 271:18(1996), pp. 10611-10615.

Characterization of an alternatively spliced G(M2) activator protein, G(M2A) protein - An activator protein which stimulates the enzymatic hydrolysis of N-acetylneuraminic acid, but not N-acetylgalactosamine, from G(M2)

S. Sonnino;
1996

Abstract

G(M2) activator protein is a protein cofactor which stimulates the enzymatic hydrolysis of both GalNAc and NeuAc from G(M2). We have previously isolated two cDNA clones, G(M2) activator cDNA and G(M2A) cDNA, for human G(M2) activator protein (Nagarajan, S., Chen, H.-C., Li, S.-C., Li, Y.-T., and Lockyer, J. M. (1992) Biochem. J. 282, 807-813). G(M2A) mRNA is an RNA alternative splicing product that contains exons 1, 2, 3, and intron 3 of the genomic DNA sequence of G(M2) activator protein (Klima, H., Tanaka, A., Schnabel, D., Nakano, T., Schroder, M., Suzuki, K., and Sandhoff, K. (1991) FEES Left. 289, 260-264). G(M2A) cDNA encodes a protein (G(M2A) protein) containing 1-109 of the 160 amino acids of human G(M2) activator protein, plus a tripeptide (VST) encoded by intron 3 at the COOH terminus. Thus, G(M2A) protein can be regarded as a form (truncated version) of G(M2) activator protein. We have expressed G(M2A) cDNA in Escherichia coli using pT7-7 as the vector. The recombinant G(M2A) protein was purified to an electrophoretically homogeneous form and was found to stimulate the hydrolysis of NeuAc from G(M2) by clostridial sialidase, but not the hydrolysis of GalNAc from G(M2) by beta-hexosaminidase A. Like G(M2) activator protein, G(M2A) protein also specifically recognized the terminal G(M2) epitope in GalNAc-GD1a and stimulated the hydrolysis of only the external NeuAc from this ganglioside by clostridial sialidase. These results enabled us to discern the enzymatic hydrolyses of GalNAc and NeuAc from the G(M2) epitope and established that the NeuAc recognition domain of G(M2) activator protein is located within amino acids 1-109. The presence of G(M2A) mRNA in human tissues and the selective stimulation of NeuAc hydrolysis by G(M2A) protein indicate that this activator protein may be involved in the catabolism of G(M2) through the asialo-G(M2) pathway.
Settore BIO/10 - Biochimica
THE JOURNAL OF BIOLOGICAL CHEMISTRY
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/188825
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