Endocytosis of N-acetylglucosamine-containing glycoproteins by rat fibroblasts expressing a single species of chicken liver glycoprotein receptor.
A cDNA clone for the chicken liver receptor which mediates endocytosis of glycoproteins containing terminal N-acetylglucosamine has been isolated and sequenced, confirming the previously obtained amino acid sequence of this protein (which is also known as the chicken hepatic lectin). This cDNA was introduced into Rat-1 fibroblasts and expressed using the promotor in the long terminal repeat of Moloney murine leukemia virus. Cells expressing chicken receptor were identified by screening with antireceptor antibodies followed by fluorescein-conjugated second antibodies. Receptor expressed in these cells was indistinguishable on gel electrophoresis from receptor isolated from liver. Three clonally isolated lines were examined for their ability to bind agalacto-alpha 1-acid glycoproteins at 0 degrees C and to take up and degrade this ligand at 37 degrees C. The receptor number (50,000/cell), affinity for ligand (35 nM), and uptake rate (5 molecules ligand/surface receptor/h) are similar to those previously observed for chicken hepatocytes, and for the uptake of asialoglycoproteins by rat hepatocytes and hepatoma cells. These findings indicate that the chicken receptor correctly traverses the endocytic pathway in a rat cell even though the cytoplasmic domain of this protein shows no primary structural homology with the corresponding portion of the rat liver receptor or with receptors found in fibroblasts.
Mellow
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Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York 10032.
Halberg
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Drickamer
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D000117:Acetylglucosamine / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002645:Chickens; D004247:DNA / Q000032:analysis; D004705:Endocytosis; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005944:Glucosamine / Q000031:analogs & derivatives*; D006023:Glycoproteins / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D008858:Microscopy, Phase-Contrast; D008969:Molecular Sequence Data; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011971:Receptors, Immunologic / Q000378:metabolism*; D014162:Transfection
D006023:Glycoproteins; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; D012333:RNA, Messenger; D011971:Receptors, Immunologic; C032193:glycoprotein receptor GPIb-IX; D004247:DNA; D005944:Glucosamine; D000117:Acetylglucosamine