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2002 | 49 | 1 | 273-283
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Adhesive properties of carcinoembryonic antigen glycoforms expressed in glycosylation-deficient Chinese hamster ovary cell lines.

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Carcinoembryonic antigen (CEA) is an oncofoetal cell surface glycoprotein that serves as an important tumour marker for colorectal and some other carcinomas. Its immunoglobulin-like structure places CEA within the immunoglobulin superfamily. CEA functions in several biological roles including homotypic and heterotypic (with other CEA family members) cell adhesion. Cell-cell interaction can be modulated by different factors, e.g., post-translational modifications such as glycosylation. The purpose of this study was to examine whether changes in carbohydrate composition of CEA oligosaccharides can influence homotypic (CEA-CEA) interactions. In order to modulate glycosylation of CEA we used two different glycosylation mutants of Chinese hamster ovary (CHO) cells, Lec2 and Lec8. Lec2 cells should produce CEA with nonsialylated N-glycans, while Lec8 cells should yield more truncated sugar structures than Lec2. Parental CHO (Pro5) cells and the glycosylation deficient mutants were stably transfected with CEA cDNA. All three CEA glycoforms, tested in a solid-phase cell adhesion assay, showed an ability to mediate CEA-dependent cell adhesion, and no qualitative differences in the adhesion between the glycoforms were observed. Thus, it may be assumed that carbohydrates do not play a role in homotypic adhesion, and the interactions between CEA molecules depend solely on the polypeptide structure.

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  • Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
  • Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
  • Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
  • Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
  • Beauchemin, N., Benchimol, S., Cournoyer, D., Fuks, A. & Stanners, C.P. (1987) Isolation and characterization of full-length functional cDNA clones for human carcinoembryonic antigen. Mol. Cell. Biol. 7, 3221-3230.
  • Benchimol, S., Fuks, A., Jothy, S., Beauchemin, N., Shirota, K. & Stanners, C.P. (1989) Carcinoembryonic antigen, a human tumor marker, functions as an intercellular adhesion molecule. Cell 57, 327-334.
  • Boehm, M.K., Mayans, M.O., Thornton, J.D., Begent, R.H.J., Keep, P.A. & Perkins, S.J. (1996) Extended glycoprotein structure of the seven domains in human carcinoembryonic antigen by X-ray and neutron solution scattering and an automated curve fitting procedure: Implications for cellular adhesion. J. Mol. Biol. 259, 718-736.
  • Chandrasekaran, E.V., Davila, M., Nixon, D.V., Goldfarb, M. & Mendicino, J. (1983) Isolation and structures of the oligosaccharide units of carcinoembryonic antigen. J. Biol. Chem. 258, 7213-7222.
  • Charbonneau, J. & Stanners, C.P. (1999) Role of carbohydrate structures in CEA-mediated intercellular adhesion. Cell Adhes. Commun. 7, 233-244.
  • Chu, T.M., Reynoso, G. & Hansen, H.J. (1972) Demonstration of carcinoembryonic antigen in normal human plasma. Nature 238, 152-153.
  • Coligan, J.E., Pritchard, D.G., Schnute, W.C., Jr. & Todd, C.W. (1976) Methylation analysis of the carbohydrate portion of carcinoembryonic antigen. Cancer Res. 36, 1915-1917.
  • Deutscher, S.L., Nuwayhid, N., Stanley, P., Briles, E.I. & Hirschberg, C.B. (1984) Translocation across Golgi vesicle membranes: A CHO glycosylation mutant deficient in CMP-sialic acid transport. Cell 39, 295-299.
  • Deutscher, S.L. & Hirschberg, C.B. (1986) Mechanism of galactosylation in the Golgi apparatus. J. Biol. Chem. 261, 96-100.
  • Diamond, M.S., Staunton, D.E., Marlin, S.D. & Springer, T.A. (1991) Binding of the integrin Mac-1 (CD11b/CD18) to the third immunoglobulin-like domain of ICAM-1 (CD54) and its regulation by glycosylation. Cell 65, 961-971.
  • Eidelman, F.J., Fuks, A., DeMarte, L., Taheri, M. & Stanners, C.P. (1993) Human carcinoembryonic antigen, an intercellular adhesion molecule, blocks fusion and differentiation of rat myoblasts. J. Cell Biol. 123, 467-475.
  • Feizi, J. (1994) Evidence for carbohydrate-mediated interactions between the neural cell adhesion molecules NCAM and L1. Trends Biochem. Sci. 19, 233-234.
  • Fieger, C.B., Emig-Vollmer, S., Petri, T., Grafe, M., Gohlke, M., Debus, N., Semmler, W., Tauber, R. & Volz, B. (2000) The adhesion properties of recombinant soluble L-selectin are modulated by its glycosylation. Biochim. Biophys. Acta 1524, 75-85.
  • Fritsche, R. & Mach, J.P. (1977) Isolation and characterization of carcinoembryonic antigen (CEA) extracted from normal human colon mucosa. Immunochemistry 14, 119-127.
  • Fukushima, K., Ohkura, T., Kanai, M., Kuroki, M., Matsuoka, Y., Kobata, A. & Yamashita, K. (1995) Carbohydrate structures of a normal counterpart of the carcinoembryonic antigen produced by colon epithelial cells of normal adults. Glycobiology 5, 105-115.
  • Garcia, M., Seigner, C., Bastid, C., Choux, R., Payan, M.J. & Reggio, H. (1991) Carcinoembryonic antigen has a different molecular weight in normal colon and in cancer cells due to N-glycosylation differences. Cancer Res. 51, 5679-5686.
  • Gold, P. & Freedman, S.O. (1965) Specific carcinoembryonic antigens of the human digestive system. J. Exp. Med. 122, 467-485.
  • Graham, F.L. & Van der Eb, A.J. (1973) A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52, 456-467.
  • Hammarström, S., Engvall, E., Johansson, B.G., Svensson, S., Sundblad, G. & Goldstein, I.J. (1975) Nature of the tumor-associated determinant(s) of carcinoembryonic antigen. Proc. Natl. Acad. Sci. U.S.A. 72, 1528-1532.
  • Hefta, S.A., Hefta, L.J.F., Lee, T.D., Paxton, R.J. & Shively, J.E. (1988) Carcinoembryonic antigen is anchored to membranes by covalent attachment to a glycosylphosphatidylinositol moiety: Identification of the ethanolamine linkage site. Proc. Natl. Acad. Sci. U.S.A. 85, 4648- 4652.
  • Hoffman, S. & Edelman, G.M. (1983) Kinetics of homophilic binding by E and A forms of the neural cell adhesion molecule. Proc. Natl. Acad. Sci. U.S.A. 80, 5762-5766.
  • Horstkorte, R., Lee, H.Y., Lucka, L., Danker, K., Mantey, L. & Reutter, W. (2001) Biochemical engineering of the side chain of sialic acids increases the biological stability of the highly sialylated cell adhesion molecule CEACAM1. Biochem. Biophys. Res. Commun. 283, 31-35.
  • Jessup, J.M., Ishii, S., Mitzoi, T., Edmiston, K.H. & Shoji, Y. (1999) Carcinoembryonic antigen facilitates experimental metastasis through a mechanism that does not involve adhesion to liver cells. Clin. Exp. Metastasis 17, 481-488.
  • Jothy, S., Yuan, S.Y. & Shirota, K. (1993) Transcription of carcinoembryonic antigen in normal colon and colon carcinoma. Am. J. Pathol. 143, 250-257.
  • Kaplan, B.E., Hefta, L.J., Blake II, R.C., Swiderek, K.M. & Shively, J.E. (1998) Solid-phase synthesis and characterization of carcinoembryonic antigen (CEA) domains. J. Peptide Res. 52, 249-260.
  • Krop-Watorek, A., Oikawa, S., Oyama, Y. & Nakazato, H. (1998) Oligomerization of N-terminal domain of carcinoembryonic antigen (CEA), expressed in Escherichia coli. Biochem. Biophys. Res. Commun. 242, 79-83.
  • Krop-Watorek, A., Laskowska, A., Salwa, J., Klopocki, A.G., Grunert, F. & Ugorski, M. (1999) CEA-related proteins on human urothelial cell lines of different transformation grades. Cancer Lett. 139, 15-22.
  • Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685.
  • Lowry, O.H., Rosebrough, N.J., Farr, A.I. & Randall, R.I. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265-275.
  • Munk, K., Pritzer, E., Kretzschmar, E., Gutte, B., Garten, W. & Klenk, H.D. (1992) Carbohydrate masking of an antigenic epitope of influenza virus haemagglutinin independent of oligosaccharide size. Glycobiology 2, 233-240.
  • Nap, M., Mollgard, K., Burtin, P. & Fleuren, G.J. (1988) Immunohistochemistry of carcino-embryonic antigen in the embryo, fetus and adult. Tumour Biol. 9, 145-153.
  • Oikawa, S., Nakazato, H. & Kosaki, G. (1987a) Primary structure of human carcinoembryonic antigen (CEA) deduced from cDNA sequence. Biochem. Biophys. Res. Commun. 142, 511-518.
  • Oikawa, S., Imajo, S., Noguchi, T., Kosaki, G. & Nakazato, H. (1987b) The carcinoembryonic antigen (CEA) contains multiple immunoglobulin-like domains. Biochem. Biophys. Res. Commun. 144, 634-642.
  • Oikawa, S., Inuzuka, C., Kuroki, Mo., Arakawa, F., Matsuoka, Y., Kosaki, G. & Nakazato, H. (1991) A specific heterotypic cell adhesion activity between members of carcinoembryonic antigen family, W272 and NCA, is mediated by N-domains. J. Biol. Chem. 266, 7995-8001.
  • Ono, M., Handa, K., Withers, D.A. & Hakomori, S. (2000) Glycosylation effect on membrane domain (GEM) involved in cell adhesion and motility: A preliminary note on functional α3, α5-CD82 glycosylation complex in ldlD 14 cells. Biochem. Biophys. Res. Commun. 279, 744-750.
  • Paxton, R.J., Mooser, G., Pande, H., Lee, T.D. & Shively, J.E. (1987) Sequence analysis of carcinoembryonic antigen: Identification of glycosylation sites and homology with the immunoglobulins supergene family. Proc. Natl. Acad. Sci. U.S.A. 84, 920-924.
  • Rudd, P.M., Wormald, M.R., Stanfield, R.L., Huang, M., Mattsson, N., Speir, J.A., DiGennaro, J.A., Fetrow, J.S., Dwek, R.A. & Wilson, I.A. (1999) Roles for glycosylation of cell surface receptors involved in cellular immune recognition. J. Mol. Biol. 293, 351-366.
  • Schrewe, H., Thompson, J., Bona, M., Hefta, L.J.F., Maruya, A., Hassauer, M., Shively, J.E., von Kleist, S. & Zimmermann, W. (1990) Cloning of the complete gene for carcinoembryonic antigen: Analysis of its promoter indicates a region conveying cell type-specific expression. Mol. Cell. Biol. 10, 2738-2748.
  • Taheri, M., Saragovi, U., Fuks, A., Makkerh, J., Mort, J. & Stanners, C.P. (2000) Self recognition in the Ig superfamily - Identification of precise subdomains in carcinoembryonic antigen required for intercellular adhesion. J. Biol. Chem. 275, 26935-26943.
  • Takami, N., Misumi, Y., Kuroki, M., Matsuoka, Y. & Ikehara, Y. (1988) Evidence for carboxyl-terminal processing and glycolipid-anchoring of human carcinoembryonic antigen. J. Biol. Chem. 263, 12716-12720.
  • Towbin, H., Staehelin, T. & Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc. Natl. Acad. Sci. U.S.A. 76, 4350-4354.
  • Von Kleist, S., Winkler, J., Migule, I. & Böhm, N. (1986) Carcinoembryonic antigen (CEA) in early embryogenesis: A study of the first trimestre of gestation. Anticancer Res. 6, 1265-1272.
  • Wagner, H.E., Toth, C.A., Steele, G.D. & Thomas, P. (1992) Metastatic potential of human colon cancer cell lines: Relationship to cellular differentiation and carcinoembryonic antigen production. Clin. Exp. Metastasis 10, 25-31.
  • Wojciechowicz, D.C., Park, P.Y., Datta, R.V. & Paty, P.B. (2000) CEA is the major PHA-L-reactive glycoprotein in colon carcinoma cell lines and tumors: Relationship between K-ras activation and beta 1-6 branching of N-linked carbohydrate on CEA. Biochem. Biophys. Res. Commun. 273, 147-153.
  • Wyss, D.F., Choi, J.S., Li, J., Knoppers, M.H., Willis, K.J., Arulanandum, A.R.N., Smolyar, A., Reinherz, E.L. & Wagner, G. (1995) Conformation and function of the N-linked glycan in the adhesion domain of human CD2. Science 269, 1273-1278.
  • Yamashita, K., Totani, K., Kuroki, M., Matsuoka, Y., Ueda, I. & Kobata, A. (1987) Structural studies of the carbohydrate moieties of carcinoembryonic antigens. Cancer Res. 47, 3451-3459.
  • Yamashita, K., Totani, K., Iwaki, Y., Kuroki, M., Matsuoka, Y., Endo, T. & Kobata, A. (1989) Carbohydrate structures of nonspecific cross- reacting antigen-2, a glycoprotein purified from meconium as an antigen cross-reacting with anticarcinoembryonic antigen antibody. J. Biol. Chem. 264, 17873-17881.
  • Zhou, H., Fuks, A., Alcaraz, G., Bolling, T.J. & Stanners, C.P. (1993) Homophilic adhesion between Ig superfamily carcinoembryonic antigen molecules involves double reciprocal bonds. J. Cell Biol. 122, 951-960.
  • Zimmermann, W., Ortlieb, B., Friedrich, R. & von Kleist, S (1987) Isolation and characterization of cDNA clones encoding the human carcinoembryonic antigen reveal a highly conserved repeating structure. Proc. Natl. Acad. Sci. U.S.A. 84, 2960-2964.
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