Role of carbohydrate structures in CEA-mediated intercellular adhesion.
Human carcinoembryonic antigen (CEA) is a member of a family of cell surface glycoproteins representing a subset of the immunoglobulin superfamily and is a major tumor marker. CEA has been demonstrated to function in vitro, at least, as a homotypic intercellular adhesion molecule. CEA can also inhibit the differentiation of several different cell types and contribute to tumorigenesis, an activity that requires CEA-CEA interactions. Post-translational modifications that could modulate CEA-CEA binding are therefore of interest. CEA is heavily glycosylated with 28 consensus sites for the addition of asparagine-linked carbohydrate structures, leading to a molecule with a bottle brush-like structure. In order to modulate the glycosylation of CEA, we transfected the functional cDNA of CEA into Chinese hamster ovary (CHO) mutant cells, Lec1, Lec2, and Lec8, which are deficient in enzymes responsible for various steps in the glycosylation processing pathway. Aggregation assays of cells in suspension were performed with stable CEA transfectants of these cell lines and showed that all of the aberrant CEA glycoforms could still mediate adhesion. In addition, the specificity of adhesion of these glycoforms was unchanged, as shown by homotypic and heterotypic adhesion assays between the transfectants. Lec1 and Lec2 transfectants did, however, show an increased speed and final extent of aggregation, which is consistent with models in which sugar structures interfere with binding through protein domains. Lec8 transfectants, on the other hand, with more truncated sugar structures than Lec2, showed less aggregation than wild type (WT) transfectants. We therefore conclude that carbohydrates do not determine the adhesion property of CEA or its specificity, in spite of the unusually high degree of glycosylation; they do, however, modulate the strength of adhesion.
Charbonneau
J
J
McGill Cancer Centre, Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
Stanners
C P
CP
D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry*; D002272:Carcinoembryonic Antigen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion; D002449:Cell Aggregation; D006224:Cricetinae; D018076:DNA, Complementary / Q000378:metabolism; D005434:Flow Cytometry; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D013997:Time Factors; D014162:Transfection
D002241:Carbohydrates; D002272:Carcinoembryonic Antigen; D018076:DNA, Complementary