A monoclonal antibody identifies a novel GPI-anchored glycoprotein involved in epithelial intercellular adhesion.
BD31 mAb, raised against a line of gastric carcinoma cells, reacts with intercellular boundaries of human transformed cells originating from carcinomas or sarcomas growing in epithelial-like clusters as well as in primary cultures of epithelial and endothelial cells. BD31 also reacts with intercellular rims of normal and transformed epithelial tissues and is particularly abundant in glands and fast-growing epithelia but absent in nervous and muscle tissues as well as in blood and in mesenchyme-derived cells. Confocal analysis indicates that BD31 is located in dots at cell-cell contacts but not in basal and apical domains of cultured and in situ epithelial cells. mAb BD31 precipitates a 100 kDa protein from cells labeled with [35S]methionine or [3H]glucosamine as well as from 125I-surface-labeled cells. This glycoprotein resists to trypsin in the presence of Ca2+, releases an 80 kDa fragment in the medium and does not react with antibodies to the conserved cytodomain of known cadherins or, specifically, to the ectodomain of E-cadherin in western blotting; moreover, the lack of cadherin cytodomain and protein removal by phosphoinositide-specific phospholipase indicate its membrane anchoring by a glycosyl-phosphatidylinositol (GPI) moiety. BD31 IgGs do not impair cell-matrix adhesion but induce inhibition of Ca(2+)-dependent aggregation, loss of cell-cell adhesion, scattering of confluent cells and appearance of migratory cell phenotypes in a dose- and time-dependent manner. This novel GPI-anchored glycoprotein may regulate intercellular adhesion by a mechanism involving membrane-associated phospholipases.
Rabino
M
M
Department of Biomedical Sciences and Human Oncology, University of Torino, Italy.
Trusolino
L
L
Prat
M
M
Cremona
O
O
Savoia
P
P
Marchisio
P C
PC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000912:Antibodies, Neoplasm / Q000276:immunology*; D002277:Carcinoma / Q000276:immunology; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000276:immunology* / Q000502:physiology; D002478:Cells, Cultured; D004847:Epithelial Cells; D004848:Epithelium / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D017261:Glycosylphosphatidylinositols; D006801:Humans; D007124:Immunoenzyme Techniques; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000502:physiology; D051379:Mice; D018613:Microscopy, Confocal; D009363:Neoplasm Proteins / Q000276:immunology; D013274:Stomach Neoplasms / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D015815:Cell Adhesion Molecules; D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins