Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Transferase other than ribonuclease
Reexamination Certificate
2001-05-10
2004-10-05
Rao, Manjunath (Department: 1652)
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Transferase other than ribonuclease
C435S069100, C435S183000, C435S252300, C435S320100, C435S333000, C435S348000, C435S349000, C435S354000, C435S358000, C536S023200, C536S023400, C536S023500, C536S024310, C536S024330, C530S350000
Reexamination Certificate
active
06800468
ABSTRACT:
1. FIELD OF THE INVENTION
The present invention relates generally to the biosynthesis of glycans found as free oligosaccharides or covalently bound to proteins and glycosphingolipids. This invention is more particularly related to a family of nucleic acids encoding UDP-D-galactose:&bgr;N-acetylglucosamine &bgr;1,3-galactosyltransferases (&bgr;3Gal-transferases), which add galactose to the hydroxy group at carbon 3 of 2-acetamido-2-deoxy-D-glucose (GlcNAc). This invention is more particularly related to a gene encoding the fifth member of the family of &bgr;3Gal-transferases, termed &bgr;3Gal-T5, probes to the DNA encoding P3Gal-T5, DNA constructs comprising DNA encoding &bgr;3Gal-T5, recombinant plasmids and recombinant methods for producing &bgr;3Gal-T5, recombinant methods for stably transfecting cells for expression of &bgr;3Gal-T5, and methods for indication of DNA polymorphism in patients.
2. BACKGROUND OF THE INVENTION
A family of UDP-galactose; &bgr;-N-acety -glucosamine &bgr;1-3galactosyl-transferases (&bgr;3Gal-T's) was recently identified (Amado, M., Almeida, R., Carneiro, F., et al. A family of human &bgr;3-gal actosyltransferases: characterisation of four members of a UDP-galactose &bgr;-N-acetylglucosamine/&bgr;-N-acetylgalactosamine &bgr;1,3-Galactosyltransferase family.
J. Biol. Chem.
273:12770-12778, 1998; Kolbinger, F., Streiff, M. B. and Katopodis, A. G. Cloning of a human UDP-galactose:2-acetamido-2-deoxy-D-glucose &bgr;3-galactosyltransferase catalysing the formation of type 1 chains.
J. Biol. Chem.
273:433-440, 1998; Hennett, T., Dinter, A., Kuhnert, P., Mattu, T. S., Rudd, P. M. and Berger, E. G. Genomic cloning and expression of three murine UDP-galactose: &bgr;-N-acetylglucosamine &bgr;1,3-galactosyltransferase genes.
J. Biol. Chem.
273:58-65, 1998; Miyaki, H., Fukumoto, S., Okada, M., Hasegawa, T. and Furukawa, K. Expression cloning of rat cDNA encoding UDP-galactose G(D2) &bgr;1,3 galactosyltransferase that determines the expression of G(D1b)/G(M 1)G(A1).
J. Biol. Chem.
272:24794-24799, 1997). Three genes within this family, &bgr;3Gal-T1, -T2, and -T3, encode &bgr;3gal actosyltransferases that form the Gal&bgr;1-3GlcNAc linkage. The type 1chain Gal&bgr;1-3GlcNAc sequence is found in both N- and O-linked oligosaccharides of glycoproteins and in lactoseries glycosphingolipids, where it is the counterpart of type 2 Gal&bgr;1-4GlcNAc poly-N-acetyllactosainine structures (Kobata. A. Structures and functions of the sugar chains of glycoproteins.
Eur J Biochem
209:483-501, 1992.). Type 1 chain structures are found mainly in endodermally derived epithelia, whereas the type 2 chains are found in ecto- and mesodermally derived cells including erythrocytes (Oriol, R., Le Pendu, J. and Mollicone, R. Genetics of ABO, H, Lewis, X and related antigens.
Vox Sanguinis
51:161-171, 1986; Clausen, H. and Hakomori, S. ABH and related histo-blood group antigens; immunochemical differences in carrier isotypes and their distribution.
Vox Sanguinis
56:1-20, 1989). Normal gastro-intestinal epithelia express mainly type 1 chain glycoconjugates, while type 2 chain structures are predominantly expressed in tumors (Hakomori, S. Aberrant glycosylation in tumors and tumor-associated carbohydrate antigens. Tumor malignancy defined by aberrant glycosylation and sphingo(glyco)lipid metabolism.
Advances in Cancer Research
52:257-331, 1989; Hakomori, S. Tumor malignancy defined by aberrant glycosylation and sphingo(glyco)lipid metabolism.
Cancer Res
56:5309-5318, 1996). It is of considerable interest to define the gene(s) responsible for formation of these core structures in normal and malignant epithelia. Several characteristics of the three previously described &bgr;3Gal-Ts capable of forming type 1 chain structures suggest that these are not the major enzyme(s) involved in type 1 chains synthesis in epithelia: (i) Northern analysis indicates that &bgr;3Gal-T1 and -T2 are exclusively expressed in brain (Amado, M., Almeida, R., Carneiro, F., et al. A family of human &bgr;3-galactosyltransferases: characterisation of four members of a UDP-galactose &bgr;-N-acetylglucosamine/&bgr;-N-acetylgalactosamine &bgr;1,3-Galactosyltransferase family.
J. Biol. Chem.
273:12770-12778, 1998; Kolbinger, F., Streiff, M. B. and Ktopodis, A. G. Cloning of a human UDP-galactose:2-acetamido-2-deoxy-D-glucose &bgr;3-galactosyltransferase catalysing the formation of type 1 chains. J. Biol. Chem. 273:433-440, 1998; Hennett, T., Dinter, A., Kubnert, P., Mattu, T. S., Rudd, P. M. and Berger, E. G. Genomic cloning and expression of three murine UDP-galactose: &bgr;-N-acetylglucosamine/&bgr;1,3-galactosyltransferase genes.
J. Biol. Chem.
273:58-65, 1998); (ii) although &bgr;3Gal-T3 has a wider expression pattern it is not detected in several tissues including colon and it is weakly expressed in gastric mucosa (Amado, M., Almeida, R., Carneiro, F., et al. A family of human &bgr;3-galactosyltransferases: characterisation of four members of a UDP-galactose &bgr;-N-acetylglucosamine/&bgr;-N-acetylgalactosamine &bgr;1,3-Galactosyltransferase family.
J. Biol. Chem.
273:12770-12778, 1998; Kolbinger, F., Streiff, M. B. and Ktopodis, A G. Cloning of a human T.JDP-galactose:2-acetamido-2-deoxy-D-glucose &bgr;3-galactosyltransferase catalysing the formation of type 1 chains.
J. Biol. Chem.
273:433-440, 1998); (iii) the kinetic properties of recombinant enzymes are not consistent with those reported for &bgr;Gal-T activities in epithelia (Sheares, B. T., Lau, J. T. and Carlson, D. M. Biosynthesis of galactosyl-beta &bgr;1,3-N-acetylgiucosamine.
J. Biol. Chem.
257:599-602, 1982; Holmes, E. H. Characterization and membrane organization of beta 1-3 and beta 1-4 galactosyltransferases from human colonic adenocarcinoma cell lines Cob 205 and SW403: basis for preferential synthesis of type 1 chain lacto-series carbohydrate structures.
Arch Biochem Biophys
270:630-646, 1989); and (iv) the acceptor substrate specificities of &bgr;3Gal-T1, -T2, or -T3 do not include the mucin-type core 3 structure (Amado, M., Almeida, R., Cameiro, F., et al. A family of human &bgr;3-galactosyltransferases: characterisation of four members of a UDP-galactose &bgr;-N-acetylglucosamine/&bgr;-N-acetylgalactosamine &bgr;1,3-Galactosyltransferase family.
J. Biol. Chem.
273:12770-12778, 1998; Hennett, T., Dinter, A., Kuhnert, P., Mattu, T. S., Rudd, P. M. and Berger, E. G. Genomic cloning and expression of three murine UDP-galactose: &bgr;3-N-acetylglucosamine &bgr;1,3-galactosyltransferase genes.
J. Biol. Chem.
273:58-65, 1998), which was previously found to be a highly efficient substrate for &bgr;3Gal-T activity isolated from porcine trachea (Sheares, B. T. and Carison, D. M. Characterization of UDP-galactose:2-acetamido-2-deoxy-D-glucose 3 beta-galactosyltransferase from pig trachea.
J. Biol. Chem.
25 8:9893-9898, 1983).
Access to additional existing &bgr;GlcNAc &bgr;3Gal-transferase genes encoding &bgr;3Gal-transferases with better kinetic properties than &bgr;3Gal-T1, -T2, and -T3 would allow production of more efficient enzymes for use in galactosylation of oligosaccharides, glycoproteins, and glycosphingolipids. Such enzymes could be used, for example, in pharmaceutical or other commercial applications that require synthetic galactosylation of these or other substrates that are not or poorly acted upon by &bgr;3Gal-T1, -T2, and -T3, in order to produce appropriately glycosylated glycoconjugates having particular enzymatic, immunogenic, or other biological and/or physical properties.
Consequently, there exists a need in the art for additional isolated UDP-galactose: &bgr;3-N-acetyl-glucosamine &bgr;1-3-Galactosyltransferases having unique, specific properties and the primary structure of the genes encoding these enzymes. The present invention meets this need, and further presents other related advantages, as described in detail below.
3. SUMMARY OF THE INVENTION
The present invention provides isolated nucleic acids encoding human UDP-galactose: &bgr;3-N-acetylglucosamine &bgr;1,3-galactosyltransferase (&bgr;3Ga
Amado Margarida
Clausen Henrik
Morgan & Lewis & Bockius, LLP
Rao Manjunath
LandOfFree
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