Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Transferase other than ribonuclease
Patent
1996-08-19
1998-12-29
Patterson, Jr., Charles L.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Transferase other than ribonuclease
435 697, C12N 910
Patent
active
058540423
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a sugar-chain synthetase and a DNA encoding the enzyme. More specifically, the present invention relates to an N-acetylgalactosamine .alpha. 2,6-sialyltransferase (GalNAc .alpha. 2,6-sialyltransferase) and a DNA encoding the enzyme. The enzyme is useful as medicaments having inhibitory activities against tumor metastases and viral infection, and as agents for introducing a sialic acid moieties into drugs to increase their biological activity.
The present invention further relates to a process for producing the sugar-chain synthetase. More specifically, the present invention relates to a process for expressing sialyltransferases in microorganisms to obtain the sialyltransferases in large quantities.
2. Decription of Related Art
Sialic acids play an important role in a variety of biological processes, like cell-cell communication, cell-substrate interaction, adhesion. It has been known that various kinds of distinguishable cell surface sialic acids exist which change in a regulated manner during development, differentiation, and oncogenic transformation.
Sialic acids occur at the terminal positions of the carbohydrate groups of glycoproteins and glycolipids, and they are enzymatically introduced from CMP-Sia to these positions in a post translational process. For example, three linkage patterns, Sia .alpha. 2,6Gal, Sia .alpha. 2,3Gal and Sia .alpha. 2,6GalNAc are commonly found in glycoproteins (Hakomori, S., Ann. Rev. Biochem., 50, pp.733-764, 1981), and two, Sia .alpha. 2,3Gal and Sia .alpha. 2,8Sia, occur frequently in gangliosides (Fishman, P., and Brady, R. O., Science, 194, pp.906-915, 1976).
The enzymes responsible for such enzymatic introduction of sialic acid (sialic acid transfer) as mentioned above are glycosyltransferases called sialyltransferases. It has been known that at least 12 different sialyltransferases are required to synthesize all known sialyloligosaccharide structures (Broquet, P. et al., Int. J. Biochem., 23, 385-389, 1991; and Weinstein, J. et al., J. Biol. Chem., 262, 17735-17743, 1987). Among these enzymes, five sialyltransferases have been purified so far, and it has been known that they exhibit strict specificity for acceptor substrate (Sadler, J. et al., J. Bio. Chem., 254, pp.4434-4443, 1979; Weinstein, J. et al., J. Biol. Chem., 257, pp.13835-13844, 1982; Rearick, J. et al., J. Biol. Chem., 254, pp.4444-4451, 1979; and Joqiasse, D. H. et al., J. Biol. Chem., 260, 4941-4951, 1985).
As for cDNAs encoding the aforementioned sialyltransferases, cDNAs encoding Gal .beta. 1,4GlcNAc .alpha. 2,6-sialyltransferase (Gal .beta. 4GlcNAc-.alpha. 6ST) have been cloned from various organs including liver (Weinstein, J. et al., J. Biol. Chem., 262, pp.17735-17743, 1987; Grundmann U. et al., Nucleic Acids Res. 18, 667, 1990; Bast, B. et al., J. Cell. Biol., 116, pp.423-435, 1992; and Hamamoto, T. et al., Bioorg. and Medic. Chem., 1, pp.141-145, 1993). Furthermore, cDNAs encoding Gal .beta. 1,3GalNAc .alpha. 2,3-sialyltransferase (Gal .beta. 3GalNAc-.alpha. 3ST) (Gillespie, W. et al., J. Biol. Chem., 267, pp.21004-21010, 1992: Japanese Patent Unexamined Publication No. 5-504678/1993; and Lee, Y. et al., Eur. J. Biochem, 216, 377-385, 1993); Gal .beta. 1,3(4) GlcNAc .alpha. 2,3-sialyltransferase (Gal .beta. 3(4)GlcNAc-.alpha. 3ST) (Wen, D. X et al., J. Biol. Chem., 267, 21011-21019,1992; and Kitagawa, H. et al., Biochem. Biophys. Res. Commun. 194, 375); and Gal .beta. 1,3GalNAc/Gal .beta. 1,4GlcNAc .alpha. 2,3-sialyltransferase (Sasaki, K. et al., J. Biol. Chem., 268, 22782-22787, 1993) have also been cloned.
With respect to GalNAc .alpha. 2,6-sialyltransferase, the isolation of this enzyme has been reported (Hakomori, S., Ann. Rev. Biochem., 50, 733-764, 1981). However, the enzyme has not been purified so as to be characterized as a single identifiable substance, and accordingly, the enzyme has not been practically used because of insufficient reaction specificity, stability, and quantitative availability.
REFERENCES:
J. Weinstein et al. "Primary Structure of .beta.-Galactoside .alpha.2,6-Sialytransferase", The Journal of Biological Chemistry, vol. 262, No. 36, pp. 17735-17743, Dec. 25, 1987.
D. Wen et al. "Primary Structure of Gal.beta.1,3(4)GlcNAc .alpha.2,3-Sialytransferase Determined by Mass Spectrometry Sequence Analysis and Molecular Cloning", The Journal of Biological Chemistry, vol. 267, No. 29, pp. 21011-21019, Oct. 15, 1992.
W. Gillespie et al. "Cloning and Expression of the Gal.beta.1,3GalNAc .alpha.2,3-Sialytransferase", The Journal of Biological Chemistry, vol. 267, No. 29, pp. 21004-21010, Oct. 15, 1992.
S. Hakomori, "Glycosphingolipids in Cellular Interaction, Differentiation, and Oncogenesis.sup.1 ", Ann. Rev. Biochem., vol. 50, pp. 733-764, 1981.
Blumenfeld, O. O., et. al. (1992) Blood 80(9), 2388-2395.
Gross, H. J., et. al. (1989) Biochemistry 28, 7386-7392.
Gross, H. J., et. al. (1988) Eur. J. Biochem. 177, 583-589.
Higa, H. H., et. al. (1985) J. Biol. Chem. 260(15),8839-8849.
Kurosawa, N, et. al. (1994) J. Biol. Chem. 269(2), 1402-1409.
Kurosawa, N, et. al. (1994) J. Biol. Chem. 269(29), 19048-19053.
J. Sadler et al., "Purification to Homogeneity of a .beta.-Galactoside .alpha.2 .fwdarw. 3 Sialytransferase and Partial Purification of an .alpha.-N-Acetylgalactosaminide .alpha.2.fwdarw.6 Sialytransferase from Porcine Submaxillary Glands", The Journal of Biological Chemistry, vol. 254, No. 11, Jun. 10, 1979, pp. 4434-4443.
J. Sadler et al., "Purification to Homogeneity and Enzymatic Characterization of an .alpha.-N-Acetylgalactosaminide .alpha.2 .fwdarw. 6 Sialyltransferase from Porcine Submaxillary Glands", The Journal of Biological Chemistry, vol. 254, No. 13, Jul. 10, 1979, pp. 5934-5941.
Hamamoto Toshiro
Kojima Naoya
Kurosawa Nobuyuki
Lee Young-Choon
Nakaoka Takashi
Patterson Jr. Charles L.
The Institute of Physical and Chemical Research
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