Compositions and methods for producing sialyltransferases

Details Compositions and methods for producing sialyltransferases Compositions and methods for producing sialyltransferases

1995-07-12

1999-01-12

Wax, Robert A.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Transferase other than ribonuclease

435 691, 4352523, 4353201, 4351723, 536 232, 530350, 935 48, C12N 910, C12N 120, C12P 2106, C07H 2104

Patent

active

058587519

DESCRIPTION:

BRIEF SUMMARY
BACKGROUND OF THE INVENTION

This invention relates to the sialyltransferase gene family, a group of glycosyltransferases responsible for the terminal sialylation of carbohydrate groups of glycoproteins, glycolipids and oligosaccharides which contain a conserved region of homology in the catalytic domain. Members of the sialyltransferase gene family comprise Gal.beta.1,3GalNAc .alpha.2,3 sialyltransferase and Gal1,3(4)GlcNAc .alpha.2,3 sialyltransferase. The invention further relates to novel forms and compositions thereof and particularly to the means and methods for the identification and production of members of the sialyltransferase gene family to homogeneity in significant useful quantities. This invention also relates to preparation of isolated deoxyribonucleic acid (DNA) coding for the production of sialyltransferases; to methods of obtaining DNA molecules which code for sialyltransferases; to the expression of human and mammalian sialyltransferases utilizing such DNA, as well as to novel compounds, including novel nucleic acids encoding sialyltransferases or fragments thereof. This invention is also directed to sialyltransferase derivatives, particularly derivatives lacking cytoplasmic and/or transmembrane portions of the protein, and their production by recombinant DNA techniques.
Sialyltransferases are a family of enzymes that catalyze the transfer of sialic acid (SA) to terminal portions on the carbohydrate groups of glycolipids and oligosaccharides in the general reaction:


Cytididine 5 monophosphate-sialic acid
(CMP-SA)+HO-acceptor.fwdarw.CMP+SA-O-Acceptor
(Beyer, T. A. et al., Adv. Enzynol., 52:23-175 (1981)). Sialyltransferases are found primarily in the Golgi apparatus of cells where they participate in post-translational glycosylation pathways. (Fleischer, B. J., Cell Biol., 89:246-255 (1981)). They are also found in body fluids, such as breast milk, colostrum and blood. At least 10-12 different sialyltransferases are required to synthesize all the sialyloligossacharide sequences known. Four sialyltransferases have been purified. (Weinstein, J. et al., J. Biol. Chem., 257:13835-13844 (1982); Miagi, T. and Tsuiki, S., Eur. J. Biochem., 125:253-261 (1982); and Joziasse, D. H. et al., J. Biol. Chem., 260:4941-4951 (1985)). More specifically, a Gal.beta.1,4GlcNAc .alpha.2-6 sialyltransferase and a Gal.beta.1,3(4) GlcNAc .alpha.2-3 sialyltransferase have been purified from rat liver membranes (Weinstein et al., ibid.).
Other glycosyltransferases have been isolated as soluble enzymes in serum, milk or colostrum including sialyl-, fucosyl-, galactosyl-, N-acetyl-gucosaminyl-, and N-acetylgalactosaminyltransferases (Beyer et al., ibid.). Bovine and human.beta.-N-acetylglucosamide.beta.1,4-galactosyltransferase has been isolated (Narimatsu, H. et al., Proc. Nat Acad. Sci. U.S.A.., 83:4720-4724 (1986); Shaper, N. L. et al., Proc. Nat. Acad. Sci. U.S.A., 83:1573-1577 (1986); Appert, H. E. et al., Biochem. Biophys. Res. Common, 139:163-168 (1986); and, Humphreys-Beyer, M. G. et al., Proc. Nat. Acad. Sci. U.S.A., 83:8918-8922 (1986). These purified glycosyltransferases differ in size which may be due to the removal of portions of the protein not essential for activity, such as the membrane spanning domains.
Comparison of the deduced amino acid sequences of the cDNA clones encoding the glycosyltransferases including galactosyltransferases, sialyltransferase, fucosyltransferase and N-acetylgalactosaminyl-transferase, reveals that these enzymes have virtually no sequence homology. Some insight into how this family of glycosyltransferases might be structurally related has come from recent analysis of the primary structures of cloned sialyltransferases (Weinstein, J. et al., ibid.). However, they all have a short NH.sub.2 -terminal cytoplasmic tail, a 16-20 amino acid signal-anchor domain, and an extended stem region which is followed by the large COOH-terminal catalytic domain Weinstein, J. et al., J. Biol. Chem., 262:17735-17743 (1987); Paulson, J. C. et al., J. Biol. Chem., 264:17615-17618 (1989). Sign

REFERENCES:
patent: 5032519 (1991-07-01), Paulson et al.
patent: 5384249 (1995-01-01), Sasaki et al.
Gillespie et al. (Oct. 15, 1992). J. Biol. Chem. 267(29): 21004-21010.
Weinstein et al. (Nov. 25, 1982). J. Biol. Chem. 257(22) : 13835-13844.
Kitagawa et al. (Jul. 15, 1993). Biochem. Biophys. Res. Comm. 194(1) : 375-382.
Sadler et al. (Jun. 10, 1979). J. Biol. Chem. 254(11) : 4434-4443.
Joziasse et al. (Apr. 25, 1985). J. Biol. Chem. 260(8): 4941-4961.
Mathews et al. (1967). J. Biol. Chem. (Jun. 5, 1987). 262(16): 7537-7545.
Lee et al. (Mar. 11, 1988). Science 239: 1288-1291.

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