Biochemical and immunological characterization of desmoplakins I and II, the major polypeptides of the desmosomal plaque.
Epithelial cells contain complexes of cytokeratin filaments (tonofilaments) with specific domains of the plasma membrane that appear as symmetric junctions, i.e. desmosomes, or as asymmetric hemi-desmosomes. These regions of filament-membrane-attachment are characterized by 14 to 20 nm thick dense plaques (desmosomal plaque). In isolated desmosome-tonofilament complexes or other desmosomal fractions from various stratified squamous epithelia (e.g. bovine muzzle epidermis and tongue mucosa) desmosomal plaque structures are recognized and show a relatively high resistance to various extraction buffers and detergents. Such fractions enriched in desmosomal plaque material are also enriched in two prominent polypeptide bands of apparent molecular weights 250,000 (desmoplakin I) and 215,000 (desmoplakin II) which appear, on two-dimensional gel electrophoresis, as two distinct polypeptides isoelectric near neutral pH. These two polypeptides are present in almost equimolar amounts and each of them appears as a series of isoelectric variants, including some labeled by [32P]phosphate in tissue slices. The two desmoplakin polypeptides are closely related as shown by tryptic peptide map analysis and are different from keratin-like proteins and other major polypeptides of desmosome-rich fractions. Guinea pig antibodies raised against desmoplakins and specific for these proteins do not cross-react with other desmosomal antigen(s) or constituents of other types of junctions. Using desmoplakin antibodies we have identified desmoplakins as the major constituents of the desmosomal plaques present in epithelial and myocardiac cells of diverse species. The significance of this group of cell type-specific membrane-associated cytoskeletal proteins and their possible cytoskeletal functions are discussed.
D000818:Animals; D000918:Antibody Specificity; D002417:Cattle; D003429:Cross Reactions; D003598:Cytoskeletal Proteins; D051180:Desmoplakins; D003896:Desmosomes / Q000032:analysis* / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology; D008854:Microscopy, Electron; D008970:Molecular Weight; D010455:Peptides / Q000276:immunology
D003598:Cytoskeletal Proteins; D051180:Desmoplakins; D008565:Membrane Proteins; D010455:Peptides