Desmoglein II-derived glycopeptides in human epidermis.
Antisera raised against a major 78 kD glycopeptide from pig epidermis were used to identify desmoglein II-derived glycopeptides in the conA-binding material isolated from human epidermis. In whole CaCl2-separated epidermis the antiserum recognized conA-binding components with apparent Mr of 115, 100, 82, 68, 50, 48, and 46 kD. The 82, 68, 48, and 46 kD immunoreactive bands were present in normal stratum corneum and plantar callus. Psoriatic scales contained significantly more of the 82 kD components and less of the 48 and 46 kD bands. Psoriatic scales also contained a major 50 kD conA-binding component unrelated to keratins or desmoglein II. Proteolytic peptide mapping showed that the major immunoreactive bands in normal stratum corneum and plantar callus were also chemically related. The 82 to 46 kD immunoreactive glycopeptides in plantar callus coincided with the major coomassie blue stained bands and were homogeneous on two-dimensional gels suggesting that this tissue may be a valuable source of human desmoglein II-derived glycopeptides. An antiserum directed against the electrophoretically co-purified 48/46 kD glycopeptides from plantar callus recognized the 82 to 46 kD bands in immunoblotting. In indirect immunofluorescence of frozen skin sections this antiserum stained the surface of epidermal cells in the spinous and granular layers of the tissue. In immunogold labeling of paraformaldehyde-fixed skin sections affinity-purified antibodies stained intact desmosomes in spinous and granular cells and desmosomal remnants in the stratum corneum. The results are consistent with our hypothesis that desmoglein II undergoes limited cleavage to stable fragments during terminal differentiation. Proteolytic degradation appears to be incomplete in psoriatic epidermis.
King
I A
IA
Dermatology Research Group, MRC Clinical Research Centre, Harrow, Middlesex U.K.
Wood
M J
MJ
Fryer
P R
PR
D003208:Concanavalin A; D003598:Cytoskeletal Proteins; D051301:Desmoglein 2; D051182:Desmogleins; D051180:Desmoplakins; D015180:Electrophoresis, Gel, Two-Dimensional; D004817:Epidermis / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006020:Glycopeptides / Q000032:analysis*; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins / Q000032:analysis*; D010449:Peptide Mapping
D003598:Cytoskeletal Proteins; D051301:Desmoglein 2; D051182:Desmogleins; D051180:Desmoplakins; D006020:Glycopeptides; D008562:Membrane Glycoproteins; D003208:Concanavalin A