title: Steric and not structure-specific factors dictate the endocytic mechanism of glycosylphosphatidylinositol-anchored proteins.
abstract: 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.
authors.affiliation: Department of Biochemistry, McGill University, Montreal, Quebec H3G1Y6, Canada.
authors.forename: Pinkesh
authors.initials: P
authors.lastname: Bhagatji
authors.forename: Rania
authors.initials: R
authors.lastname: Leventis
authors.forename: Jonathan
authors.initials: J
authors.lastname: Comeau
authors.forename: Mohammad
authors.initials: M
authors.lastname: Refaei
authors.forename: John R
authors.initials: JR
authors.lastname: Silvius
mesh_terms: 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
chemical_list: 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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