Retention of prominin in microvilli reveals distinct cholesterol-based lipid micro-domains in the apical plasma membrane.
Membrane cholesterol-sphingolipid 'rafts', which are characterized by their insolubility in the non-ionic detergent Triton X-100 in the cold, have been implicated in the sorting of certain membrane proteins, such as placental alkaline phosphatase (PLAP), to the apical plasma membrane domain of epithelial cells. Here we show that prominin, an apically sorted pentaspan membrane protein, becomes associated in the trans-Golgi network with a lipid raft that is soluble in Triton X-100 but insoluble in another non-ionic detergent, Lubrol WX. At the cell surface, prominin remains insoluble in Lubrol WX and is selectively associated with microvilli, being largely segregated from the membrane subdomains containing PLAP. Cholesterol depletion results in the loss of prominin's microvillus-specific localization but does not lead to its complete intermixing with PLAP. We propose the coexistence within a membrane domain, such as the apical plasma membrane, of different cholesterol-based lipid rafts, which underlie the generation and maintenance of membrane subdomains.
Röper
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K
Department of Neurobiology, Interdisciplinary Center of Neuroscience, University of Heidelberg, Im Neuenheimer Feld 364, D-69120 Heidelberg, Germany.
Corbeil
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D
Huttner
W B
WB
D000071916:AC133 Antigen; D000469:Alkaline Phosphatase; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D015703:Antigens, CD; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism*; D003432:Cross-Linking Reagents; D003599:Cytoskeleton / Q000378:metabolism; D003902:Detergents; D004285:Dogs; D058851:GPI-Linked Proteins; D006023:Glycoproteins; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D050356:Lipid Metabolism; D008562:Membrane Glycoproteins / Q000276:immunology / Q000378:metabolism*; D021962:Membrane Microdomains / Q000378:metabolism*; D008871:Microvilli / Q000378:metabolism; D017830:Octoxynol; D010455:Peptides; D011092:Polyethylene Glycols; D012995:Solubility; D021601:trans-Golgi Network / Q000378:metabolism
D000071916:AC133 Antigen; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D003432:Cross-Linking Reagents; D003902:Detergents; D058851:GPI-Linked Proteins; D006023:Glycoproteins; D017261:Glycosylphosphatidylinositols; D007527:Isoenzymes; D008562:Membrane Glycoproteins; D010455:Peptides; C017474:Lubrol WX; D011092:Polyethylene Glycols; D017830:Octoxynol; D002784:Cholesterol; D000469:Alkaline Phosphatase; C019839:alkaline phosphatase, placental