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

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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

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Caveolae
Smooth Muscle Myocytes
Chickens
Caveolins
Avian Gizzard
Connective Tissue Cells
Transcytosis
Heterotrimeric GTP-Binding Proteins
Glycosylphosphatidylinositols
Membranes
Monomeric GTP-Binding Proteins
Electrophoresis
Signal Transduction
Membrane Proteins
Proteins
Epithelium
Endothelial Cells
Biomarkers
Gels
Cell Membrane

ASJC Scopus subject areas

  • Cell Biology

Cite this

Chang, W. J., Ying, Y. S., Rothberg, K. G., Hooper, N. M., Turner, A. J., Gambliel, H. A., ... 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

Purification and characterization of smooth muscle cell caveolae. / Chang, Wen Jinn; Ying, Yun Shu; Rothberg, Karen G.; Hooper, Nigel M.; Turner, Anthony J.; Gambliel, Hervé A.; De Gunzburg, Jean; Mumby, Susanne M.; Gilman, Alfred G.; Anderson, Richard G W.

In: Journal of Cell Biology, Vol. 126, No. 1, 07.1994, p. 127-138.

Research output: Contribution to journalArticle

Chang, WJ, Ying, YS, Rothberg, KG, Hooper, NM, Turner, AJ, Gambliel, HA, De Gunzburg, J, Mumby, SM, Gilman, AG & Anderson, RGW 1994, 'Purification and characterization of smooth muscle cell caveolae', Journal of Cell Biology, vol. 126, no. 1, pp. 127-138. https://doi.org/10.1083/jcb.126.1.127
Chang WJ, Ying YS, Rothberg KG, Hooper NM, Turner AJ, Gambliel HA et al. Purification and characterization of smooth muscle cell caveolae. Journal of Cell Biology. 1994 Jul;126(1):127-138. https://doi.org/10.1083/jcb.126.1.127
Chang, Wen Jinn ; Ying, Yun Shu ; Rothberg, Karen G. ; Hooper, Nigel M. ; Turner, Anthony J. ; Gambliel, Hervé A. ; De Gunzburg, Jean ; Mumby, Susanne M. ; Gilman, Alfred G. ; Anderson, Richard G W. / Purification and characterization of smooth muscle cell caveolae. In: Journal of Cell Biology. 1994 ; Vol. 126, No. 1. pp. 127-138.
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