Phorbol ester induces increased expression, altered glycosylation, and reduced adhesion of K562 erythroleukemia cell fibronectin receptors.
The human multipotential hematopoietic cell line K562 expresses fibronectin receptor (FNR) subunits of 160 kDa (alpha chain) and 120 kDa (beta chain). Treatment with 12-O-tetradecanoylphorbol 13-acetate (TPA) led to reduced binding of K562 to immobilized fibronectin (FN), although treated cells expressed 10-fold more cell surface FNR than untreated cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis confirmed this and showed altered electrophoretic mobilities of FNR subunits from TPA-treated cells. TPA treatment affected N-linked glycosylation, as tunicamycin treatment of K562 cells abolished differences in FNR mobility. Sialidase treatment of FNR immunoprecipitates minimized and sialidase treatment of intact cells eliminated these mobility differences between subunits from control and TPA-treated cells. Reduced sialylation of FNR from TPA-treated cells was further demonstrated by chromatography with bead-coupled lectins and by the greater negative charge of untreated K562 FNR subunits in two-dimensional isoelectric focusing-polyacrylamide gel electrophoresis. A relationship between reduced FNR sialylation and reduced FN binding was suggested by adhesion assays of sialidase-treated K562 which showed that desialylation of cell surface FNR was associated with decreased cell adhesion. Thus, TPA treatment reduces the function, increases the expression, and alters the structure of K562 FNR, and these changes appear to involve FNR sialylation.
Symington
B E
BE
Fred Hutchinson Cancer Research Center, Division of Basic, Seattle, Washington 98104.
Symington
F W
FW
Rohrschneider
L R
LR
D002448:Cell Adhesion / Q000187:drug effects*; D002460:Cell Line; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism* / Q000473:pathology; D009439:Neuraminidase; D010703:Phorbol Esters / Q000494:pharmacology; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic / Q000187:drug effects* / Q000302:isolation & purification / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology
D005353:Fibronectins; D010703:Phorbol Esters; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic; C008876:phorbol-12,13-didecanoate; D009439:Neuraminidase; D013755:Tetradecanoylphorbol Acetate