Expression of distinct desmocollin isoforms in human epidermis.
Previous evidence suggested the presence of two distinct desmocollin isoforms in human epidermis. These isoforms have now been distinguished at the protein level using monoclonal and polyclonal antibodies against N-terminal fragments of desmosomal glycoprotein (DG) IV/V isolated from plantar callus and antibodies against a fusion protein containing the extracellular domain of DGII/III. Immune blotting of glycoprotein fractions from whole epidermis, plantar callus, psoriatic scales and cultured keratinocytes showed that intact DGIV/V and its proteolytic fragments consistently migrated faster than DGII/III during SDS-PAGE. The apparent Mr difference between the two isoforms was in the range 2-5 kD. DGIV/V was the predominant species in epidermal tissue but was much less prominent in cultured cells by immune-blotting and immune precipitation. This is consistent with the differentiation-related expression of desmocollins revealed by immunofluorescence. DGIV/V was strongly expressed in the upper spinous/granular layer of the epidermis whereas DGII/III was more prominent in the basal layers of the tissue. The DGIV/V monoclonal (LH50) recognized an N-terminal, Ca(++)-sensitive epitope, because its staining of unfixed epidermal tissue was markedly influenced by Ca++ levels. Ca++ inhibition was observed at concentrations as low as 50 microM, suggesting its possible physiologic significance. Ca++ inhibition of LH50 binding was also observed in an enzyme-linked immunosorbent assay system using denatured glycoproteins although higher concentrations were required. It remains to be seen whether direct effects of Ca++ on desmocollin conformation are involved in the regulation of keratinization by extracellular Ca++.
King
I A
IA
Dermatology Research Centre, Harrow, London, U.K.
Tabiowo
A
A
Purkis
P
P
Leigh
I
I
Magee
A I
AI
D000911:Antibodies, Monoclonal / Q000187:drug effects; D002146:Bony Callus / Q000473:pathology; D002118:Calcium / Q000032:analysis / Q000494:pharmacology; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000032:analysis*; D051187:Desmocollins; D051180:Desmoplakins; D003896:Desmosomes / Q000737:chemistry; D004817:Epidermis / Q000737:chemistry; D006801:Humans; D015151:Immunoblotting; D007536:Isomerism; D015603:Keratinocytes / Q000166:cytology; D008297:Male; D011565:Psoriasis / Q000473:pathology; D012867:Skin / Q000737:chemistry*; D013194:Staining and Labeling
D000911:Antibodies, Monoclonal; D003598:Cytoskeletal Proteins; D051187:Desmocollins; D051180:Desmoplakins; D002118:Calcium