Dynamin-mediated internalization of caveolae

John R Henley, E. W A Krueger, Barbara J. Oswald, Mark A Mc Niven

Research output: Contribution to journalArticle

576 Citations (Scopus)

Abstract

The dynamins comprise an expanding family of ubiquitously expressed 100- kD GTPases that have been implicated in severing clathrin-coated pits during receptor-mediated endocytosis. Currently, it is unclear whether the different dynamin isoforms perform redundant functions or participate in distinct endocytic processes. To define the function of dynamin II in mammalian epithelial cells, we have generated and characterized peptide-specific antibodies to domains that either are unique to this isoform or conserved within the dynamin family. When microinjected into cultured hepatocytes these affinity-purified antibodies inhibited clathrin-mediated endocytosis and induced the formation of long plasmalemmal invaginations with attached clathrin-coated pits. In addition, clusters of distinct, nonclathrin-coated, flask-shaped invaginations resembling caveolae accumulated at the plasma membrane of antibody-injected cells. In support of this, caveola-mediated endocytosis of labeled cholera toxin B was inhibited in antibody-injected hepatocytes. Using immunoisolation techniques an anti-dynamin antibody isolated caveolar membranes directly from a hepatocyte postnuclear membrane fraction. Finally, double label immunofluorescence microscopy revealed a striking colocalization between dynamin and the caveolar coat protein caveolin. Thus, functional in vivo studies as well as ultrastructural and biochemical analyses indicate that dynamin mediates both clathrin-dependent endocytosis and the internalization of caveolae in mammalian cells.

Original languageEnglish (US)
Pages (from-to)85-99
Number of pages15
JournalJournal of Cell Biology
Volume141
Issue number1
DOIs
StatePublished - Apr 6 1998

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Dynamins
Caveolae
Clathrin
Endocytosis
Hepatocytes
Antibodies
Protein Isoforms
Dynamin II
Caveolins
Antibody Affinity
Membranes
Cholera Toxin
GTP Phosphohydrolases
Capsid Proteins
Fluorescence Microscopy
Anti-Idiotypic Antibodies
Epithelial Cells
Cell Membrane
Peptides

ASJC Scopus subject areas

  • Cell Biology

Cite this

Dynamin-mediated internalization of caveolae. / Henley, John R; Krueger, E. W A; Oswald, Barbara J.; Mc Niven, Mark A.

In: Journal of Cell Biology, Vol. 141, No. 1, 06.04.1998, p. 85-99.

Research output: Contribution to journalArticle

Henley, John R ; Krueger, E. W A ; Oswald, Barbara J. ; Mc Niven, Mark A. / Dynamin-mediated internalization of caveolae. In: Journal of Cell Biology. 1998 ; Vol. 141, No. 1. pp. 85-99.
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