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VIP21-caveolin, a membrane protein constituent of the caveolar coat, oligomerizes in vivo and in vitro

S Monier, RG Parton, F Vogel, J Behlke, A Henske and TV Kurzchalia

Department of Cell Biology, Max-Delbruck Centre for Molecular Medicine, Berlin-Buch, Germany.

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.

Volume 6, Issue 7, pp. 911-927, 07/01/1995
Copyright © 1995 by The American Society for Cell Biology




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R. Luetterforst, E. Stang, N. Zorzi, A. Carozzi, M. Way, and R. G. Parton
Molecular Characterization of Caveolin Association with the Golgi Complex: Identification of a Cis-Golgi Targeting Domain in the Caveolin Molecule
J. Cell Biol., June 28, 1999; 145(7): 1443 - 1459.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
K. Das, R. Y. Lewis, P. E. Scherer, and M. P. Lisanti
The Membrane-spanning Domains of Caveolins-1 and -2 Mediate the Formation of Caveolin Hetero-oligomers. IMPLICATIONS FOR THE ASSEMBLY OF CAVEOLAE MEMBRANES IN VIVO
J. Biol. Chem., June 25, 1999; 274(26): 18721 - 18728.
[Abstract] [Full Text] [PDF]


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Circ. Res.Home page
Y.-Y. Zhao, O. Feron, C. Dessy, X. Han, M. A. Marchionni, and R. A. Kelly
Neuregulin Signaling in the Heart : Dynamic Targeting of erbB4 to Caveolar Microdomains in Cardiac Myocytes
Circ. Res., June 25, 1999; 84(12): 1380 - 1387.
[Abstract] [Full Text] [PDF]


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Mol. Biol. CellHome page
R. Nomura and T. Fujimoto
Tyrosine-phosphorylated Caveolin-1: Immunolocalization and Molecular Characterization
Mol. Biol. Cell, April 1, 1999; 10(4): 975 - 986.
[Abstract] [Full Text]


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J. Biol. Chem.Home page
K. A. Melkonian, A. G. Ostermeyer, J. Z. Chen, M. G. Roth, and D. A. Brown
Role of Lipid Modifications in Targeting Proteins to Detergent-resistant Membrane Rafts. MANY RAFT PROTEINS ARE ACYLATED, WHILE FEW ARE PRENYLATED
J. Biol. Chem., February 5, 1999; 274(6): 3910 - 3917.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
V. Rizzo, D. P. McIntosh, P. Oh, and J. E. Schnitzer
In Situ Flow Activates Endothelial Nitric Oxide Synthase in Luminal Caveolae of Endothelium with Rapid Caveolin Dissociation and Calmodulin Association
J. Biol. Chem., December 25, 1998; 273(52): 34724 - 34729.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
V. Rizzo, A. Sung, P. Oh, and J. E. Schnitzer
Rapid Mechanotransduction in Situ at the Luminal Cell Surface of Vascular Endothelium and Its Caveolae
J. Biol. Chem., October 9, 1998; 273(41): 26323 - 26329.
[Abstract] [Full Text] [PDF]


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J. Lipid Res.Home page
F. J. Field, E. Born, S. Murthy, and S. N. Mathur
Caveolin is present in intestinal cells: role in cholesterol trafficking?
J. Lipid Res., October 1, 1998; 39(10): 1938 - 1950.
[Abstract] [Full Text]


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J. Biol. Chem.Home page
H. Al-Hasani, C. S. Hinck, and S. W. Cushman
Endocytosis of the Glucose Transporter GLUT4 Is Mediated by the GTPase Dynamin
J. Biol. Chem., July 10, 1998; 273(28): 17504 - 17510.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
L. Snyers, E. Umlauf, and R. Prohaska
Oligomeric Nature of the Integral Membrane Protein Stomatin
J. Biol. Chem., July 3, 1998; 273(27): 17221 - 17226.
[Abstract] [Full Text] [PDF]


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JCBHome page
P. Oh, D. P. McIntosh, and J. E. Schnitzer
Dynamin at the Neck of Caveolae Mediates Their Budding to Form Transport Vesicles by GTP-driven Fission from the Plasma Membrane of Endothelium
J. Cell Biol., April 6, 1998; 141(1): 101 - 114.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
T. Okamoto, A. Schlegel, P. E. Scherer, and M. P. Lisanti
Caveolins, a Family of Scaffolding Proteins for Organizing "Preassembled Signaling Complexes" at the Plasma Membrane
J. Biol. Chem., March 6, 1998; 273(10): 5419 - 5422.
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JCBHome page
S. Esser, K. Wolburg, H. Wolburg, G. Breier, T. Kurzchalia, and W. Risau
Vascular Endothelial Growth Factor Induces Endothelial Fenestrations In Vitro
J. Cell Biol., February 23, 1998; 140(4): 947 - 959.
[Abstract] [Full Text] [PDF]


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JCBHome page
P. Scheiffele, P. Verkade, A.M. Fra, H. Virta, K. Simons, and E. Ikonen
Caveolin-1 and -2 in the Exocytic Pathway of MDCK Cells
J. Cell Biol., February 23, 1998; 140(4): 795 - 806.
[Abstract] [Full Text] [PDF]


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JCBHome page
C. Lipardi, R. Mora, V. Colomer, S. Paladino, L. Nitsch, E. Rodriguez-Boulan, and C. Zurzolo
Caveolin Transfection Results in Caveolae Formation but Not Apical Sorting of Glycosylphosphatidylinositol (GPI)-anchored Proteins in Epithelial Cells
J. Cell Biol., February 9, 1998; 140(3): 617 - 626.
[Abstract] [Full Text] [PDF]


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J. Lipid Res.Home page
D. Hailstones, L. S. Sleer, R. G. Parton, and K. K. Stanley
Regulation of caveolin and caveolae by cholesterol in MDCK cells
J. Lipid Res., February 1, 1998; 39(2): 369 - 379.
[Abstract] [Full Text]


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J. Cell Sci.Home page
U Vogel, K Sandvig, and B van Deurs
Expression of caveolin-1 and polarized formation of invaginated caveolae in Caco-2 and MDCK II cells
J. Cell Sci., January 3, 1998; 111(6): 825 - 832.
[Abstract] [PDF]


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J. Biol. Chem.Home page
J. Couet, M. Sargiacomo, and M. P. Lisanti
Interaction of a Receptor Tyrosine Kinase, EGF-R, with Caveolins. CAVEOLIN BINDING NEGATIVELY REGULATES TYROSINE AND SERINE/THREONINE KINASE ACTIVITIES
J. Biol. Chem., November 28, 1997; 272(48): 30429 - 30438.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
P. E. Scherer, R. Y. Lewis, D. Volonte, J. A. Engelman, F. Galbiati, J. Couet, D. S. Kohtz, E. van Donselaar, P. Peters, and M. P. Lisanti
Cell-type and Tissue-specific Expression of Caveolin-2. CAVEOLINS 1 AND 2 CO-LOCALIZE AND FORM A STABLE HETERO-OLIGOMERIC COMPLEX IN VIVO
J. Biol. Chem., November 14, 1997; 272(46): 29337 - 29346.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
P. E. Scherer and M. P. Lisanti
Association of Phosphofructokinase-M with Caveolin-3 in Differentiated Skeletal Myotubes. DYNAMIC REGULATION BY EXTRACELLULAR GLUCOSE AND INTRACELLULAR METABOLITES
J. Biol. Chem., August 15, 1997; 272(33): 20698 - 20705.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
R. Puertollano, S. Li, M. P. Lisanti, and M. A. Alonso
Recombinant Expression of the MAL Proteolipid, a Component of Glycolipid-enriched Membrane Microdomains, Induces the Formation of Vesicular Structures in Insect Cells
J. Biol. Chem., July 18, 1997; 272(29): 18311 - 18315.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
O. Feron, T. W. Smith, T. Michel, and R. A. Kelly
Dynamic Targeting of the Agonist-stimulated m2 Muscarinic Acetylcholine Receptor to Caveolae in Cardiac Myocytes
J. Biol. Chem., July 11, 1997; 272(28): 17744 - 17748.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
J. Liu, P. Oh, T. Horner, R. A. Rogers, and J. E. Schnitzer
Organized Endothelial Cell Surface Signal Transduction in Caveolae Distinct from Glycosylphosphatidylinositol-anchored Protein Microdomains
J. Biol. Chem., March 14, 1997; 272(11): 7211 - 7222.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
J. Couet, S. Li, T. Okamoto, T. Ikezu, and M. P. Lisanti
Identification of Peptide and Protein Ligands for the Caveolin-scaffolding Domain. IMPLICATIONS FOR THE INTERACTION OF CAVEOLIN WITH CAVEOLAE-ASSOCIATED PROTEINS
J. Biol. Chem., March 7, 1997; 272(10): 6525 - 6533.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
K. S. Song, Z. Tang, S. Li, and M. P. Lisanti
Mutational Analysis of the Properties of Caveolin-1. A NOVEL ROLE FOR THE C-TERMINAL DOMAIN IN MEDIATING HOMO-TYPIC CAVEOLIN-CAVEOLIN INTERACTIONS
J. Biol. Chem., February 14, 1997; 272(7): 4398 - 4403.
[Abstract] [Full Text] [PDF]


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J. Histochem. Cytochem.Home page
T. Fujimoto and K. Fujimoto
Metal Sandwich Method to Quick-freeze Monolayer Cultured Cells for Freeze-fracture
J. Histochem. Cytochem., February 1, 1997; 45(4): 595 - 598.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
Z. Tang, T. Okamoto, P. Boontrakulpoontawee, T. Katada, A. J. Otsuka, and M. P. Lisanti
Identification, Sequence, and Expression of an Invertebrate Caveolin Gene Family from the Nematode Caenorhabditis elegans. IMPLICATIONS FOR THE MOLECULAR EVOLUTION OF MAMMALIAN CAVEOLIN GENES
J. Biol. Chem., January 24, 1997; 272(4): 2437 - 2445.
[Abstract] [Full Text] [PDF]


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JCBHome page
R. G. Parton, M. Way, N. Zorzi, and E. Stang
Caveolin-3 Associates with Developing T-tubules during Muscle Differentiation
J. Cell Biol., January 13, 1997; 136(1): 137 - 154.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
S. Li, J. Couet, and M. P. Lisanti
Src Tyrosine Kinases, Galpha Subunits, and H-Ras Share a Common Membrane-anchored Scaffolding Protein, Caveolin. CAVEOLIN BINDING NEGATIVELY REGULATES THE AUTO-ACTIVATION OF Src TYROSINE KINASES
J. Biol. Chem., November 15, 1996; 271(46): 29182 - 29190.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
S. Li, K. S. Song, S. S. Koh, A. Kikuchi, and M. P. Lisanti
Baculovirus-based Expression of Mammalian Caveolin in Sf21 Insect Cells. A MODEL SYSTEM FOR THE BIOCHEMICAL AND MORPHOLOGICAL STUDY OF CAVEOLAE BIOGENESIS
J. Biol. Chem., November 8, 1996; 271(45): 28647 - 28654.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
K. S. Song, P. E. Scherer, Z. Tang, T. Okamoto, S. Li, M. Chafel, C. Chu, D. S. Kohtz, and M. P. Lisanti
Expression of Caveolin-3 in Skeletal, Cardiac, and Smooth Muscle Cells. CAVEOLIN-3 IS A COMPONENT OF THE SARCOLEMMA AND CO-FRACTIONATES WITH DYSTROPHIN AND DYSTROPHIN-ASSOCIATED GLYCOPROTEINS
J. Biol. Chem., June 21, 1996; 271(25): 15160 - 15165.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
K. S. Song, S. Li, T. Okamoto, L. A. Quilliam, M. Sargiacomo, and M. P. Lisanti
Co-purification and Direct Interaction of Ras with Caveolin, an Integral Membrane Protein of Caveolae Microdomains
J. Biol. Chem., April 19, 1996; 271(16): 9690 - 9697.
[Abstract] [Full Text] [PDF]




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