Extracellular domain of pemphigus vulgaris antigen (desmoglein 3) mediates weak homophilic adhesion.
Pemphigus vulgaris antigen is in the cadherin supergene family. We hypothesized that the extracellular domain of pemphigus vulgaris antigen might mediate homophilic cell adhesion because 1) the originally described cadherins (e.g., E-cadherin) mediate this type of adhesion, 2) pemphigus vulgaris antigen is localized in desmosomes that are cell adhesion junctions, and 3) autoantibodies in pemphigus vulgaris patients cause loss of cell adhesion. To test this hypothesis we used a system developed for E-cadherin that, when transfected into L cells (mouse fibroblasts), has been shown to cause aggregation. Because this aggregation requires the cytoplasmic domain of E-cadherin to bind to catenins, we made a chimeric cDNA construct that encodes the extracellular domain of pemphigus vulgaris antigen and the cytoplasmic domain of E-cadherin. Analysis by immunofluorescence and flow cytometry with pemphigus vulgaris sera indicated that the pemphigus vulgaris antigen extracellular domain of this chimeric molecule (PVEC) was expressed on the cell surface of transiently transfected cells and permanently transfected L-cell clones. Immunoprecipitation of the chimeric molecule from extracts of these clones showed that the E-cadherin cytoplasmic domain bound catenins. Surprisingly, these L-cell clones displayed only slight aggregation compared to an L-cell clone transfected with E-cadherin. This weak aggregation was, however, specific and homophilic, as determined by cell sorting of only PVEC transfectants into aggregates from mixtures of PVEC and neomycin resistance gene transfectants, one of which was labeled with a fluorescent dye. We conclude that the extracellular domain of pemphigus vulgaris antigen mediates weak homophilic adhesion and is not interchangeable in function with the extracellular domain of E-cadherin.
Amagai
M
M
Dermatology Branch, National Institutes of Health, Bethesda, Maryland 20892.
Kàrpàti
S
S
Klaus-Kovtun
V
V
Udey
M C
MC
Stanley
J R
JR
D000818:Animals; D015820:Cadherins / Q000737:chemistry*; D002448:Cell Adhesion; D002678:Chimera / Q000235:genetics; D003593:Cytoplasm / Q000737:chemistry; D018076:DNA, Complementary / Q000032:analysis; D051184:Desmoglein 3; D007739:L Cells / Q000276:immunology / Q000502:physiology; D051379:Mice; D010392:Pemphigus / Q000276:immunology; D014162:Transfection
D015820:Cadherins; D018076:DNA, Complementary; C071894:DSG3 protein, human; D051184:Desmoglein 3