The tryptic cleavage product of the mature form of the bovine desmoglein 1 ectodomain is one of the antigen moieties immunoprecipitated by all sera from symptomatic patients affected by a new variant of endemic pemphigus.
Multiple antigens are recognized by sera from patients with pemphigus foliaceus (PF). Several have been identified including keratin 59, desmocollins, envoplakin, periplakin, and desmogleins 1 and 3 (Dsg1 and Dsg3). In addition, an 80 kDa antigen was identified as the N-terminal fragment of Dsg1 using as antigen source an insoluble epidermal cell envelope preparation. However, still unsolved was the identity of the most important antigenic moiety, a 45 kDa tryptic fragment which is recognized by all sera from patients with fogo selvagem, pemphigus foliaceus, by half of pemphigus vulgaris sera and by a new variant of endemic pemphigus in E1 Bagre, Colombia that resembles Senear-Usher syndrome. Here, we report the identification of the 45 kDa conformational epitope of a soluble tryptic cleavage product from viable bovine epidermis. To elucidate the nature of this peptide, viable bovine epidermis was trypsin-digested, and glycosylated peptides were partially purified on a concanavalin A (Con-A) affinity column. This column fraction was then used as an antigen source for further immunoaffinity purification. A PF patient's serum covalently coupled to a Staphylococcus aureus protein A column was incubated with the Con-A eluted products and the immuno-isolated antigen was separated by SDS-PAGE, transferred to a membrane, and visualized with Coomassie blue, silver and amido black stains. The 45 kD band was subjected to amino acid sequence analysis revealing the sequence, EXIKFAAAXREGED, which matched the mature form of the extracellular domain of bovine Dsg1. This study confirms the biological importance of the ectodomain of Dsg1 as well as the relevance of conformational epitopes in various types of pemphigus.
Abréu-Vélez
Ana María
AM
Institute for Molecular Medicine and Genetics, Medical College of Georgia, CB 2803, 1120, 15th Street, Augusta, GA 30912-2630, USA. aavelez@mail.mcg.edu
Javier Patiño
Pablo
P
Montoya
Fernando
F
Bollag
Wendy B
WB
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis*; D015820:Cadherins / Q000032:analysis / Q000276:immunology*; D016022:Case-Control Studies; D002417:Cattle; D051183:Desmoglein 1; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D010392:Pemphigus / Q000097:blood* / Q000276:immunology*; D010446:Peptide Fragments / Q000032:analysis; D011233:Precipitin Tests
D000954:Antigens, Surface; D015820:Cadherins; D051183:Desmoglein 1; D010446:Peptide Fragments