VIP21-caveolin, a membrane protein constituent of the caveolar coat, oligomerizes in vivo and in vitro.
VIP21-caveolin is a membrane protein, proposed to be a component of the striated coat covering the cytoplasmic surface of caveolae. To investigate the biochemical composition of the caveolar coat, we used our previous observation that VIP21-caveolin is present in large complexes and insoluble in the detergents CHAPS or Triton X-114. The mild treatment of these insoluble structures with sodium dodecyl sulfate leads to the detection of high molecular mass complexes of approximately 200, 400, and 600 kDa. The 400-kDa complex purified to homogeneity from dog lung is shown to consist exclusive of the two isoforms of VIP21-caveolin. Pulse-chase experiments indicate that the oligomers form early after the protein is synthesized in the endoplasmic reticulum (ER). VIP21-caveolin does indeed insert into the ER membrane through the classical translocation machinery. Its hydrophobic domain adopts an unusual loop configuration exposing the N- and C-flanking regions to the cytoplasm. Similar high molecular mass complexes can be produced from the in vitro-synthesized VIP21-caveolin. The complex formation occurs only if VIP21-caveolin isoforms are properly inserted into the membrane; formation is cytosol-dependent and does not involve a vesicle fusion step. We propose that high molecular mass oligomers of VIP21-caveolin represent the basic units forming the caveolar coat. They are formed in the ER and later, between the ER and the plasma membrane, these oligomers could associate into larger detergent-insoluble structures.
Monier
S
S
Department of Cell Biology, Max-Delbrück Centre for Molecular Medicine, Berlin-Buch, Germany.
Parton
R G
RG
Vogel
F
F
Behlke
J
J
Henske
A
A
Kurzchalia
T V
TV
D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D051242:Caveolin 1; D022461:Caveolins; D002462:Cell Membrane / Q000737:chemistry*; D002478:Cells, Cultured; D002793:Cholic Acids; D003902:Detergents; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000378:metabolism; D007668:Kidney / Q000737:chemistry / Q000166:cytology; D008168:Lung / Q000737:chemistry / Q000166:cytology; D008561:Membrane Fusion; D008565:Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008861:Microsomes / Q000378:metabolism; D008970:Molecular Weight; D017830:Octoxynol; D011092:Polyethylene Glycols; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D012967:Sodium Dodecyl Sulfate; D012995:Solubility
D002352:Carrier Proteins; D051242:Caveolin 1; D022461:Caveolins; D002793:Cholic Acids; D003902:Detergents; D008565:Membrane Proteins; D021382:Protein Sorting Signals; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate