Glycosylation of CD45: carbohydrate processing through Golgi apparatus is required for cell surface expression and protein stability.
The importance of glycosylation on cell surface expression, protein stability and function of the CD45 phosphotyrosine phosphatase leukocyte common antigen, has been studied in the K562 erythroleukemic cell line. Cell treatment with the N-glycosylation inhibitor tunicamycin generated unglycosylated CD45 polypeptides of 130 and 140 kDa. Immunofluorescence flow cytometry and Scatchard techniques revealed that CD45 cell surface expression was decreased in a time- and dose-dependent manner upon tunicamycin incubation. Moreover, a remarkable decrease in CD45 phosphatase activity was detected in tunicamycin-treated cells, which correlated with the diminished CD45 cell surface expression. Pulse-chase biosynthetic assays demonstrated that the fate of the unglycosylated CD45 proteins underwent a late non-lysosomal degradation, since it was not prevented either by monensin, an inhibitor of Golgi transport, or by the lysosomal function inhibitor chloroquine. These results demonstrate that intact glycosylation of CD45 molecules through Golgi compartment is required for stability and proper transport towards the plasma membrane of these phosphotyrosine phosphatase proteins.
Pulido
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R
Sección de Inmunología, Hospital de la Princesa, Universidad Autónoma de Madrid, Spain.
Sánchez-Madrid
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F
D015703:Antigens, CD / Q000737:chemistry / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D006031:Glycosylation; D006649:Histocompatibility Antigens / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D017493:Leukocyte Common Antigens; D008970:Molecular Weight; D008985:Monensin / Q000494:pharmacology; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D014415:Tunicamycin / Q000494:pharmacology
D015703:Antigens, CD; D006649:Histocompatibility Antigens; D014415:Tunicamycin; D002738:Chloroquine; D008985:Monensin; D017493:Leukocyte Common Antigens