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J. Biol. Chem., Vol. 265, Issue 1, 114-125, Jan, 1990
Human transferrin receptor contains O-linked oligosaccharides
SI Do, C Enns and RD Cummings
Department of Biochemistry, University of Georgia, Athens 30602.
We have investigated the oligosaccharides in the human transferrin receptor
from three different cell lines. During our studies on the structures of
the N-linked oligosaccharides of the receptor, we discovered that the
receptor contains O-linked oligosaccharides. This report describes the
isolation and characterization of these O-linked oligosaccharides. Three
different human cell lines--K562, A431, and BeWo--were grown in media
containing either [2-3H] mannose or [6- 3H]glucosamine. The newly
synthesized and radiolabeled transferrin receptors were purified by
immunoprecipitation from cell extracts and sodium dodecyl
sulfate-polyacrylamide gel electrophoresis. The receptor was
proteolytically digested or treated directly with mild base/borohydride.
The released radiolabeled glycopeptides and oligosaccharides were separated
by a variety of chromatographic techniques, and their structures were
analyzed. The transferrin receptor from all three cell types contains
O-linked oligosaccharides that are released from peptide by mild
base/borohydride treatment. The receptor from K562 cells contains at least
one O-linked oligosaccharide having two sialic acid residues and a core
structure of the disaccharide galactose-N-acetyl-galactosamine. In
contrast, the O- linked oligosaccharides in the transferring receptors from
both A431 and BeWo cell lines are not as highly sialylated and were
identified as both the neutral disaccharide galactose-N-acetylgalactosamine
and the neutral monosaccharide N-acetylgalactosamine. In addition, the
receptors from all three cell lines contain both complex-type and high
mannose-type N-linked oligosaccharides. The complex-type chains in the
receptor from A431 cells have properties of blood group A antigens, whereas
oligosaccharides in receptors from both BeWo and K562 cells lack these
properties. These results are interesting since both A431 and BeWo cells,
but not K562 cells, are positive for blood group A antigens. Thus, our
results demonstrate that the human transferrin receptor contains O-linked
oligosaccharides and that there are differences in the structures of both
the O-linked and complex-type N- linked oligosaccharides on the receptors
synthesized by different cell types.

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Copyright © 1990 by the American Society for Biochemistry and Molecular Biology.
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