GPI membrane anchor is determinant in intracellular accumulation of apical plasma membrane proteins in the non-polarized human colon cancer cell line HT-29 18.
We have compared the intracellular localization of plasma membrane proteins anchored either with a transmembrane segment or with a glycosylphosphatidylinositol moiety to estimate the effects of membrane anchor on protein segregation in the non-polarized form of the human colon cancer cell line HT-29 18. We have monitored two endogenous proteins: the carcinoembryonic antigen, a glycosylphosphatidylinositol protein and the transmembrane protein dipeptidyl peptidase IV, and two transfected proteins: the glycosylphosphatidylinositol protein Thy-1 and an engineered transmembrane form of Thy-1. Using immunocytochemistry on ultra-thin cryosections and confocal microscopy, we detected a carcinoembryonic antigen-rich vesicular compartment, excluding classical pre-lysosomal and lysosomal markers such as mannose 6-phosphate receptor, lamp-1 and cathepsin D. This compartment, where carcinoembryonic antigen accumulated, excluded the transmembrane protein dipeptidyl peptidase IV and was reduced during the polarization of the cells. Moreover, the glycosylphosphatidylinositol form of Thy-1 also accumulated in the carcinoembryonic antigen-rich compartment whereas the transmembrane form of Thy-1 was excluded. We proposed that, in the non-polarized HT-29 18 cells, accumulation of glycosylphosphatidylinositol proteins independently of transmembrane proteins reveals different intracellular fates for proteins according to their anchor in the plasma membrane.
Nicolas
F
F
Laboratoire de Génétique et de Physiologie du Développement, UMR CNRS 9943, Faculté des Sciences de Luminy, Marseille, France.
Tiveron
M C
MC
Davoust
J
J
Reggio
H
H
D002272:Carcinoembryonic Antigen / Q000032:analysis; D002462:Cell Membrane / Q000502:physiology; D016764:Cell Polarity / Q000502:physiology; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000473:pathology; D018819:Dipeptidyl Peptidase 4 / Q000032:analysis; D017261:Glycosylphosphatidylinositols / Q000502:physiology*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D018800:Thy-1 Antigens / Q000032:analysis; D014407:Tumor Cells, Cultured
D002272:Carcinoembryonic Antigen; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D018800:Thy-1 Antigens; D018819:Dipeptidyl Peptidase 4