Purification and characterization of smooth muscle cell caveolae

Wen Jinn Chang, Yun Shu Ying, Karen G. Rothberg, Nigel M. Hooper, Anthony J. Turner, Hervé A. Gambliel, Jean De Gunzburg, Susanne M. Mumby, Alfred G. Gilman, Richard G W Anderson

Research output: Contribution to journalArticle

307 Scopus citations

Abstract

Plasmalemmal caveolae are a membrane specialization that mediates transcytosis across endothelial cells and the uptake of small molecules and ions by both epithelial and connective tissue cells. Recent findings suggest that caveolae may, in addition, be involved in signal transduction. To better understand the molecular composition of this membrane specialization, we have developed a biochemical method for purifying caveolae from chicken smooth muscle cells. Biochemical and morphological markers indicate that we can obtain ~1.5 mg of protein in the caveolae fraction from ~100 g of chicken gizzard. Gel electrophoresis shows that there are more than 30 proteins enriched in caveolae relative to the plasma membrane. Among these proteins are: caveolin, a structural molecule of the caveolae coat; multiple, glycosylphosphatidylinositol-anchored membrane proteins; both G(α) and G(β) subunits of heterotrimeric GTP-binding protein; and the Ras-related GTP- binding protein, Rap1A/B. The method we have developed will facilitate future studies on the structure and function of caveolae.

Original languageEnglish (US)
Pages (from-to)127-138
Number of pages12
JournalJournal of Cell Biology
Volume126
Issue number1
DOIs
StatePublished - Jul 1994

ASJC Scopus subject areas

  • Cell Biology

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    Chang, W. J., Ying, Y. S., Rothberg, K. G., Hooper, N. M., Turner, A. J., Gambliel, H. A., De Gunzburg, J., Mumby, S. M., Gilman, A. G., & Anderson, R. G. W. (1994). Purification and characterization of smooth muscle cell caveolae. Journal of Cell Biology, 126(1), 127-138. https://doi.org/10.1083/jcb.126.1.127