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Vol. 10, Issue 4, 961-974, April 1999
-Cyclodextrin Perturbs
Formation of Clathrin-coated Endocytic Vesicles

and
*Institute for Cancer Research, The Norwegian Radium Hospital,
Montebello, 0310 Oslo, Norway; and The importance of cholesterol for endocytosis has been investigated
in HEp-2 and other cell lines by using methyl-
Structural Cell
Biology Unit, Department of Medical Anatomy, The Panum Institute,
University of Copenhagen, DK-2200 Copenhagen N, Denmark
-cyclodextrin (M
CD)
to selectively extract cholesterol from the plasma membrane. M
CD
treatment strongly inhibited endocytosis of transferrin and EGF,
whereas endocytosis of ricin was less affected. The inhibition of
transferrin endocytosis was completely reversible. On removal of M
CD
it was restored by continued incubation of the cells even in serum-free
medium. The recovery in serum-free medium was inhibited by addition of
lovastatin, which prevents cholesterol synthesis, but endocytosis
recovered when a water-soluble form of cholesterol was added together
with lovastatin. Electron microscopical studies of M
CD-treated HEp-2
cells revealed that typical invaginated caveolae were no longer
present. Moreover, the invagination of clathrin-coated pits was
strongly inhibited, resulting in accumulation of shallow coated pits.
Quantitative immunogold labeling showed that transferrin receptors were
concentrated in coated pits to the same degree (approximately
sevenfold) after M
CD treatment as in control cells. Our results
therefore indicate that although clathrin-independent (and
caveolae-independent) endocytosis still operates after removal of
cholesterol, cholesterol is essential for the formation of
clathrin-coated endocytic vesicles.
Corresponding author. E-mail
address: ksandvig{at}radium.uio.no.
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E. N. Dedkova, X. Ji, S. L. Lipsius, and L. A. Blatter Mitochondrial calcium uptake stimulates nitric oxide production in mitochondria of bovine vascular endothelial cells Am J Physiol Cell Physiol, February 1, 2004; 286(2): C406 - C415. [Abstract] [Full Text] |
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H. Lu, T.-X. Sun, R. Bouley, K. Blackburn, M. McLaughlin, and D. Brown Inhibition of endocytosis causes phosphorylation (S256)-independent plasma membrane accumulation of AQP2 Am J Physiol Renal Physiol, February 1, 2004; 286(2): F233 - F243. [Abstract] [Full Text] [PDF] |
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B. G. Galvez, S. Matias-Roman, M. Yanez-Mo, M. Vicente-Manzanares, F. Sanchez-Madrid, and A. G. Arroyo Caveolae Are a Novel Pathway for Membrane-Type 1 Matrix Metalloproteinase Traffic in Human Endothelial Cells Mol. Biol. Cell, February 1, 2004; 15(2): 678 - 687. [Abstract] [Full Text] [PDF] |
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G. Salazar, R. Love, E. Werner, M. M. Doucette, S. Cheng, A. Levey, and V. Faundez The Zinc Transporter ZnT3 Interacts with AP-3 and It Is Preferentially Targeted to a Distinct Synaptic Vesicle Subpopulation Mol. Biol. Cell, February 1, 2004; 15(2): 575 - 587. [Abstract] [Full Text] [PDF] |
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L. H. J. Aarts, O. Roovers, A. C. Ward, and I. P. Touw Receptor activation and 2 distinct COOH-terminal motifs control G-CSF receptor distribution and internalization kinetics Blood, January 15, 2004; 103(2): 571 - 579. [Abstract] [Full Text] [PDF] |
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E. Stuven, A. Porat, F. Shimron, E. Fass, D. Kaloyanova, B. Brugger, F. T. Wieland, Z. Elazar, and J. B. Helms Intra-Golgi Protein Transport Depends on a Cholesterol Balance in the Lipid Membrane J. Biol. Chem., December 26, 2003; 278(52): 53112 - 53122. [Abstract] [Full Text] [PDF] |
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R. Hnasko and M. P. Lisanti The Biology of Caveolae: Lessons from Caveolin Knockout Mice and Implications for Human Disease Mol. Interv., December 1, 2003; 3(8): 445 - 464. [Abstract] [Full Text] [PDF] |
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S. Gummuluru, M. Rogel, L. Stamatatos, and M. Emerman Binding of Human Immunodeficiency Virus Type 1 to Immature Dendritic Cells Can Occur Independently of DC-SIGN and Mannose Binding C-Type Lectin Receptors via a Cholesterol-Dependent Pathway J. Virol., December 1, 2003; 77(23): 12865 - 12874. [Abstract] [Full Text] [PDF] |
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B. Nichols Caveosomes and endocytosis of lipid rafts J. Cell Sci., December 1, 2003; 116(23): 4707 - 4714. [Abstract] [Full Text] [PDF] |
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J. Kwik, S. Boyle, D. Fooksman, L. Margolis, M. P. Sheetz, and M. Edidin Membrane cholesterol, lateral mobility, and the phosphatidylinositol 4,5-bisphosphate-dependent organization of cell actin PNAS, November 25, 2003; 100(24): 13964 - 13969. [Abstract] [Full Text] [PDF] |
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N. B. Chauhan Membrane dynamics, cholesterol homeostasis, and Alzheimer's disease J. Lipid Res., November 1, 2003; 44(11): 2019 - 2029. [Abstract] [Full Text] |
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A. Rapacciuolo, S. Suvarna, L. Barki-Harrington, L. M. Luttrell, M. Cong, R. J. Lefkowitz, and H. A. Rockman Protein Kinase A and G Protein-coupled Receptor Kinase Phosphorylation Mediates {beta}-1 Adrenergic Receptor Endocytosis through Different Pathways J. Biol. Chem., September 12, 2003; 278(37): 35403 - 35411. [Abstract] [Full Text] [PDF] |
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C. Cobbold, J. Coventry, S. Ponnambalam, and A. P. Monaco The Menkes disease ATPase (ATP7A) is internalized via a Rac1-regulated, clathrin- and caveolae-independent pathway Hum. Mol. Genet., July 1, 2003; 12(13): 1523 - 1533. [Abstract] [Full Text] [PDF] |
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E. Elenko, T. Fischer, I. Niesman, T. Harding, T. McQuistan, M. Von Zastrow, and M. G. Farquhar Spatial Regulation of G{alpha}i Protein Signaling in Clathrin-Coated Membrane Microdomains Containing GAIP Mol. Pharmacol., July 1, 2003; 64(1): 11 - 20. [Abstract] [Full Text] [PDF] |
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L. K. Nyasae, A. L. Hubbard, and P. L. Tuma Transcytotic Efflux from Early Endosomes Is Dependent on Cholesterol and Glycosphingolipids in Polarized Hepatic Cells Mol. Biol. Cell, July 1, 2003; 14(7): 2689 - 2705. [Abstract] [Full Text] [PDF] |
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E. Coeytaux, D. Coulaud, E. Le Cam, O. Danos, and A. Kichler The Cationic Amphipathic alpha -Helix of HIV-1 Viral Protein R (Vpr) Binds to Nucleic Acids, Permeabilizes Membranes, and Efficiently Transfects Cells J. Biol. Chem., May 9, 2003; 278(20): 18110 - 18116. [Abstract] [Full Text] [PDF] |
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L. Snyers, H. Zwickl, and D. Blaas Human Rhinovirus Type 2 Is Internalized by Clathrin-Mediated Endocytosis J. Virol., May 1, 2003; 77(9): 5360 - 5369. [Abstract] [Full Text] [PDF] |
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D. Brown The ins and outs of aquaporin-2 trafficking Am J Physiol Renal Physiol, May 1, 2003; 284(5): F893 - F901. [Abstract] [Full Text] [PDF] |
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S. Shigematsu, R. T. Watson, A. H. Khan, and J. E. Pessin The Adipocyte Plasma Membrane Caveolin Functional/Structural Organization Is Necessary for the Efficient Endocytosis of GLUT4 J. Biol. Chem., March 14, 2003; 278(12): 10683 - 10690. [Abstract] [Full Text] [PDF] |
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M. J. Petris, K. Smith, J. Lee, and D. J. Thiele Copper-stimulated Endocytosis and Degradation of the Human Copper Transporter, hCtr1 J. Biol. Chem., March 7, 2003; 278(11): 9639 - 9646. [Abstract] [Full Text] [PDF] |
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K. J. D. Ashbourne Excoffon, T. Moninger, and J. Zabner The Coxsackie B Virus and Adenovirus Receptor Resides in a Distinct Membrane Microdomain J. Virol., February 15, 2003; 77(4): 2559 - 2567. [Abstract] [Full Text] [PDF] |
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