Functionally different GPI proteins are organized in different domains on the neuronal surface.
We have investigated the organization, on the plasma membrane and in detergent-insoluble membrane vesicles, of two neuronal glycosylphosphatidylinositol-anchored (GPI) proteins: Thy-1, a negative regulator of transmembrane signalling; and prion protein, whose rapid endocytosis and Cu(2+) binding suggest that it functions in metal ion uptake. Prion protein occurred on the neuronal surface at high density in domains, located primarily at the cell body, which were relatively soluble in detergent. Thy-1, although much more abundantly expressed on neurons, occurred at lower density over much of the surface of neurites (and in lower abundance at the cell body) in domains that were highly resistant to detergent solubilization. Detergent-insoluble membrane vesicles contained Thy-1 at a density similar to that on the neuronal surface. Vesicles containing each protein could be separated by immunoaffinity isolation; lectin binding showed that they were enriched in different glycoproteins. Our results demonstrate a structural diversity of the domains occupied by functionally different GPI proteins.
Madore
N
N
Molecular Neurobiology Group, New Hunt's House, EM Unit, GKT Medical and Dental School, Guy's Campus, London Bridge, London SE1 9RT, UK.
Smith
K L
KL
Graham
C H
CH
Jen
A
A
Brady
K
K
Hall
S
S
Morris
R
R
D000818:Animals; D000918:Antibody Specificity; D002458:Cell Fractionation; D002462:Cell Membrane / Q000648:ultrastructure*; D002499:Centrifugation, Density Gradient; D003902:Detergents; D017261:Glycosylphosphatidylinositols / Q000032:analysis*; D037102:Lectins; D008562:Membrane Glycoproteins / Q000032:analysis*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D016253:Microscopy, Immunoelectron; D016501:Neurites / Q000648:ultrastructure*; D009474:Neurons / Q000648:ultrastructure*; D011328:Prions / Q000032:analysis*; D051381:Rats; D012995:Solubility; D018800:Thy-1 Antigens / Q000032:analysis*
D003902:Detergents; D017261:Glycosylphosphatidylinositols; D037102:Lectins; D008562:Membrane Glycoproteins; D011328:Prions; D018800:Thy-1 Antigens