title: Induction of caveolae in the apical plasma membrane of Madin-Darby canine kidney cells.
abstract: In this paper, we have analyzed the behavior of antibody cross-linked raft-associated proteins on the surface of MDCK cells. We observed that cross-linking of membrane proteins gave different results depending on whether cross-linking occurred on the apical or basolateral plasma membrane. Whereas antibody cross-linking induced the formation of large clusters on the basolateral membrane, resembling those observed on the surface of fibroblasts (Harder, T., P. Scheiffele, P. Verkade, and K. Simons. 1998. J. Cell Biol. 929-942), only small ( approximately 100 nm) clusters formed on the apical plasma membrane. Cross-linked apical raft proteins e.g., GPI-anchored placental alkaline phosphatase (PLAP), influenza hemagglutinin, and gp114 coclustered and were internalized slowly ( approximately 10% after 60 min). Endocytosis occurred through surface invaginations that corresponded in size to caveolae and were labeled with caveolin-1 antibodies. Upon cholesterol depletion the internalization of PLAP was completely inhibited. In contrast, when a non-raft protein, the mutant LDL receptor LDLR-CT22, was cross-linked, it was excluded from the clusters of raft proteins and was rapidly internalized via clathrin-coated pits. Since caveolae are normally present on the basolateral membrane but lacking from the apical side, our data demonstrate that antibody cross-linking induced the formation of caveolae, which slowly internalized cross-linked clusters of raft-associated proteins.
authors.affiliation: European Molecular Biology Laboratory, Cell Biology and Biophysics Programme, D-69117 Heidelberg, Germany.
authors.forename: P
authors.initials: P
authors.lastname: Verkade
authors.forename: T
authors.initials: T
authors.lastname: Harder
authors.forename: F
authors.initials: F
authors.lastname: Lafont
authors.forename: K
authors.initials: K
authors.lastname: Simons
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D000906:Antibodies; D051242:Caveolin 1; D022461:Caveolins; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D016764:Cell Polarity; D002784:Cholesterol / Q000378:metabolism; D003034:Coated Pits, Cell-Membrane / Q000737:chemistry / Q000378:metabolism; D003432:Cross-Linking Reagents; D004285:Dogs; D004705:Endocytosis; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry / Q000378:metabolism; D007668:Kidney; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D008970:Molecular Weight; D009154:Mutation / Q000235:genetics; D015388:Organelles / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure; D011973:Receptors, LDL / Q000235:genetics / Q000378:metabolism
chemical_list: D000906:Antibodies; D051242:Caveolin 1; D022461:Caveolins; D003432:Cross-Linking Reagents; D006023:Glycoproteins; D017261:Glycosylphosphatidylinositols; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D008565:Membrane Proteins; D011973:Receptors, LDL; D002784:Cholesterol; D000469:Alkaline Phosphatase