Selective anchoring in the specific plasma membrane domain

A role in epithelial cell polarity

Pedro J Salas, D. E. Vega-Salas, J. Hochman, E. Rodriguez-Boulan, M. Edidin

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

We have studied the role of restrictions to lateral mobility in the segregation of proteins to apical and basolateral domains of MDCK epithelial cells. Radioimmunoassay and semiquantitative video analysis of immunofluorescence on frozen sections showed that one apical and three basolateral glycoproteins, defined by monoclonal antibodies and binding of beta-2-microglobulin, were incompletely extracted with 0.5% Triton X-100 in a buffer that preserves the cortical cytoskeleton (Fey, E.G., K.M. Wan, and S. Penman. 1984. J. Cell Biol. 98:1973-1984; Nelson, W.T. and P.J. Veshnock. 1986. J. Cell Biol. 103:1751-1766). The marker proteins were preferentially extracted from the 'incorrect' domain (i.e., the apical domain for a basolateral marker), indicating that the cytoskeletal anchoring was most effective on the 'correct' domain. The two basolateral markers were unpolarized and almost completely extractable in cells prevented from establishing cell-cell contacts by incubation in low Ca++ medium, while an apical marker was only extracted from the basal surface under the same conditions. Procedures were developed to apply fluorescent probes to either the apical or the basolateral surface of live cells grown on native collagen gels. Fluorescence recovery after photobleaching of predominantly basolateral antigens showed a large percent of cells (28-52%) with no recoverable fluorescence on the basal domain but normal fluorescence recovery on the apical surface of most cells (92-100%). Diffusion coefficients in cells with normal fluorescence recovery were in the order of 1.1 x 10-9 cm2/s in the apical domain and 0.6-0.9 x 10-9 cm2/s in the basal surface, but the difference was not significant. Te data from both techniques indicate (a) the existence of mobile and immobile protein fractions in both plasma membrane domains, and (b) that linkage to a domain specific submembrane cytoskeleton plays an important role in the maintenance of epithelial cell surface polarity.

Original languageEnglish
Pages (from-to)2363-2376
Number of pages14
JournalJournal of Cell Biology
Volume107
Issue number6 I
StatePublished - Dec 1 1988
Externally publishedYes

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Cell Polarity
Epithelial Cells
Cell Membrane
Fluorescence
Cytoskeleton
Fluorescence Recovery After Photobleaching
beta 2-Microglobulin
Proteins
Madin Darby Canine Kidney Cells
Octoxynol
Frozen Sections
Fluorescent Dyes
Radioimmunoassay
Fluorescent Antibody Technique
Glycoproteins
Buffers
Collagen
Gels
Monoclonal Antibodies
Maintenance

ASJC Scopus subject areas

  • Cell Biology

Cite this

Salas, P. J., Vega-Salas, D. E., Hochman, J., Rodriguez-Boulan, E., & Edidin, M. (1988). Selective anchoring in the specific plasma membrane domain: A role in epithelial cell polarity. Journal of Cell Biology, 107(6 I), 2363-2376.

Selective anchoring in the specific plasma membrane domain : A role in epithelial cell polarity. / Salas, Pedro J; Vega-Salas, D. E.; Hochman, J.; Rodriguez-Boulan, E.; Edidin, M.

In: Journal of Cell Biology, Vol. 107, No. 6 I, 01.12.1988, p. 2363-2376.

Research output: Contribution to journalArticle

Salas, PJ, Vega-Salas, DE, Hochman, J, Rodriguez-Boulan, E & Edidin, M 1988, 'Selective anchoring in the specific plasma membrane domain: A role in epithelial cell polarity', Journal of Cell Biology, vol. 107, no. 6 I, pp. 2363-2376.
Salas PJ, Vega-Salas DE, Hochman J, Rodriguez-Boulan E, Edidin M. Selective anchoring in the specific plasma membrane domain: A role in epithelial cell polarity. Journal of Cell Biology. 1988 Dec 1;107(6 I):2363-2376.
Salas, Pedro J ; Vega-Salas, D. E. ; Hochman, J. ; Rodriguez-Boulan, E. ; Edidin, M. / Selective anchoring in the specific plasma membrane domain : A role in epithelial cell polarity. In: Journal of Cell Biology. 1988 ; Vol. 107, No. 6 I. pp. 2363-2376.
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