Steric and not structure-specific factors dictate the endocytic mechanism of glycosylphosphatidylinositol-anchored proteins.
Diverse glycosylphosphatidylinositol (GPI)-anchored proteins enter mammalian cells via the clathrin- and dynamin-independent, Arf1-regulated GPI-enriched early endosomal compartment/clathrin-independent carrier endocytic pathway. To characterize the determinants of GPI protein targeting to this pathway, we have used fluorescence microscopic analyses to compare the internalization of artificial lipid-anchored proteins, endogenous membrane proteins, and membrane lipid markers in Chinese hamster ovary cells. Soluble proteins, anchored to cell-inserted saturated or unsaturated phosphatidylethanolamine (PE)-polyethyleneglycols (PEGs), closely resemble the GPI-anchored folate receptor but differ markedly from the transferrin receptor, membrane lipid markers, and even protein-free PE-PEGs, both in their distribution in peripheral endocytic vesicles and in the manner in which their endocytic uptake responds to manipulations of cellular Arf1 or dynamin activity. These findings suggest that the distinctive endocytic targeting of GPI proteins requires neither biospecific recognition of their GPI anchors nor affinity for ordered-lipid microdomains but is determined by a more fundamental property, the steric bulk of the lipid-anchored protein.
Bhagatji
Pinkesh
P
Department of Biochemistry, McGill University, Montreal, Quebec H3G1Y6, Canada.
Leventis
Rania
R
Comeau
Jonathan
J
Refaei
Mohammad
M
Silvius
John R
JR
D020823:ADP-Ribosylation Factor 1 / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000737:chemistry / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D034281:Dynamins / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology*; D005492:Folic Acid / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D015394:Molecular Structure; D021381:Protein Transport / Q000502:physiology; D019809:Streptavidin / Q000378:metabolism; D014168:Transferrin / Q000235:genetics / Q000378:metabolism
D015415:Biomarkers; D017261:Glycosylphosphatidylinositols; D008563:Membrane Lipids; D008565:Membrane Proteins; D014168:Transferrin; D019809:Streptavidin; D005492:Folic Acid; D020823:ADP-Ribosylation Factor 1; D034281:Dynamins
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