Characterization of a keratinocyte-specific extracellular epitope of desmoglein. Implications for desmoglein heterogeneity and function.
Despite the presumed importance of desmoglein, a 160-kDa glycoprotein, in desmosome formation and its possible involvement in certain blistering skin diseases, the precise location and function of this protein have not yet been firmly established. We describe here the characterization of a new monoclonal antibody, AE23, against an extracellular epitope of desmoglein. Both the AE23 epitope and another epitope, defined by the previously characterized DG3.4 antibody, reside on a 160-kDa human epidermal desmoglein as evidenced by their identical solubility profile, their coexistence in a 130-kDa desmoglein degradative product, their coadsorption by an AE23 immunoaffinity column, and the identical changes in the two antigens' electrophoretic mobility after air oxidation and deglycosylation. The AE23 epitope is resistant to various endoglycosidases, suggesting that sugar moieties are not involved. Characterization of several proteolytic fragments of this epidermal desmoglein enabled us to map the DG3.4 epitope to a 96-kDa intracellular domain and the AE23 epitope to an extracellular domain flanked by the plasma membrane and the distal N-glycosylation site(s). However, these two epitopes do not always coexist on the same desmoglein molecule. For example, tissue surveys showed that although the DG3.4 epitope is present in the desmogleins of all epithelial cell types, the AE23 epitope is limited to normal keratinocytes. Moreover, electron microscopic localization data indicate that whereas the DG3.4 epitope is detected in the submembranous plaques of desmosomes, the AE23 epitope is present in the intercellular space of both desmosomal and nondesmosomal areas. These results raise the possibility that there exist several biochemically closely related isoforms of desmoglein, one (AE23+/DG3.4+) restricted to epidermal desmosomes, one (AE23+/DG3.4-) uniformly distributed along the keratinocyte cell surface, and another (AE23-/DG3.4+) present in desmosomes of simple epithelia and basal cells of cultured keratinocytes. The uniform distribution of at least one desmoglein-related antigen in the intercellular space of keratinocytes coupled with the realization that different isoforms of desmogleins form a subfamily of cadherins suggest that desmoglein(s) may play a more general role in keratinocyte adhesion than previously appreciated.
Loomis
C A
CA
Department of Dermatology, Kaplan Comprehensive Cancer Center, New York University Medical School, New York 10016.
Kolega
J
J
Manabe
M
M
Sun
T T
TT
D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002462:Cell Membrane / Q000648:ultrastructure; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003598:Cytoskeletal Proteins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D051182:Desmogleins; D051180:Desmoplakins; D003896:Desmosomes / Q000648:ultrastructure*; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis* / Q000302:isolation & purification; D005455:Fluorescent Antibody Technique; D006026:Glycoside Hydrolases; D006801:Humans; D007124:Immunoenzyme Techniques; D015603:Keratinocytes / Q000166:cytology* / Q000648:ultrastructure; D008252:Macaca fascicularis; D008254:Macaca nemestrina; D016253:Microscopy, Immunoelectron; D008970:Molecular Weight
D000911:Antibodies, Monoclonal; D003598:Cytoskeletal Proteins; D051182:Desmogleins; D051180:Desmoplakins; D000939:Epitopes; D006026:Glycoside Hydrolases