Identification of a Ca2+-dependent cell-cell adhesion molecule in endothelial cells

Ronald L Heimark, M. Degner, S. M. Schwartz

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

Confluent cultures of aortic endothelial cells contain two different cell-cell adhesion mechanisms distinguished by their requirement for calcium during trypsinization and adhesion. A hybridoma clone was isolated producing a monoclonal antibody Ec6C10, which inhibits Ca2+-dependent adhesion of endothelial cells. There was no inhibition of Ca2+-independent adhesion of endothelial cells and only a minor effect on Ca2+-dependent adhesion of smooth muscle cells. Immunoblotting analysis shows that the antibody Ec6C10 recognizes a protein in endothelial but not epithelial cells with an apparent molecular weight of 135,000 in reducing conditions and 130,000 in non-reducing conditions. Monoclonal antibody Ec6C10 reacts with an antigen at the cell surface as shown by indirect immunofluorescence of confluent endothelial cells in a junctional pattern outlining the cobblestone morphology of the monolayer. Removal of extracellular calcium increased the susceptibility of the antigen recognized by antibody Ec6C10 to proteolysis by trypsin. The role of the Ca2+-dependent cell adhesion molecule in organization of the dense peripheral microfilament band in confluent endothelial cells was examined by adjusting the level of extracellular calcium to modulate cell-cell contact. Addition of the monoclonal antibody Ec6C10 at the time of the calcium switch inhibited the extent of formation of the peripheral F-actin band. These results suggest an association between cell-cell contact and the peripheral F-actin band potentially through the Ca2+-dependent CAM.

Original languageEnglish (US)
Pages (from-to)1745-1756
Number of pages12
JournalJournal of Cell Biology
Volume110
Issue number5
DOIs
StatePublished - 1990
Externally publishedYes

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Cell Adhesion Molecules
Endothelial Cells
Calcium
Monoclonal Antibodies
Actins
Antibodies
Hybridomas
Surface Antigens
Indirect Fluorescent Antibody Technique
Actin Cytoskeleton
Immunoblotting
Cell Adhesion
Trypsin
Proteolysis
Smooth Muscle Myocytes
Clone Cells
Molecular Weight
Epithelial Cells
Antigens
Proteins

ASJC Scopus subject areas

  • Cell Biology

Cite this

Identification of a Ca2+-dependent cell-cell adhesion molecule in endothelial cells. / Heimark, Ronald L; Degner, M.; Schwartz, S. M.

In: Journal of Cell Biology, Vol. 110, No. 5, 1990, p. 1745-1756.

Research output: Contribution to journalArticle

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