A mutated transferrin receptor lacking asparagine-linked glycosylation sites shows reduced functionality and an association with binding immunoglobulin protein.
The function of the transferrin receptor is to transport iron-bound transferrin into the cell. In order to function properly, this dimeric glycoprotein must be expressed on the cell surface and be able to bind transferrin. Site-directed mutagenesis was performed to abolish the three asparagine-linked glycosylation consensus sequences of the human transferrin receptor. The DNA encoding the mutated transferrin receptor was stably transfected into mouse fibroblasts. This form of the human transferrin receptor shows reduced transferrin binding, reduced intersubunit bond formation, and reduced cell surface expression, indicating that the transferrin receptor which lacks asparagine-linked glycosylation is not fully functional. In addition, the mutated form of the receptor is not processed as quickly. It shows an association with an endoplasmic reticular chaperone protein, binding immunoglobulin protein, leading to the hypothesis that the mutated transferrin receptor experiences increased retention in the endoplasmic reticulum.
Williams
A M
AM
Department of Cell Biology and Anatomy, Oregon Health Sciences University, Portland 97201-3098.
Enns
C A
CA
D001216:Asparagine / Q000737:chemistry*; D001483:Base Sequence; D002352:Carrier Proteins / Q000378:metabolism*; D003854:Deoxyribonucleotides; D004220:Disulfides / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism*; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011233:Precipitin Tests; D011990:Receptors, Transferrin / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D002352:Carrier Proteins; D003854:Deoxyribonucleotides; D004220:Disulfides; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D007136:Immunoglobulins; D018832:Molecular Chaperones; D011990:Receptors, Transferrin; D001216:Asparagine