title: The mapping of linear B-cell epitope regions in the extracellular parts of the desmoglein 1 and 3 proteins: recognition of immobilized peptides by pemphigus patients' serum autoantibodies.
abstract: Desmosomal transmembrane glycoproteins desmogleins (Dsg) 1 and 3 are targets of life-threatening autoimmune blistering disorders such as Pemphigus vulgaris (PV) and Pemphigus foliaceus (PF). In these diseases, pemphigus autoantibodies are produced against Dsg1 and Dsg3 proteins. The autoantibodies bind to these transmembrane elements leading to a loss of desmosomal cell-cell adhesion and clinically, to the presence of blisters and erosions. Identification, characterization, and detailed analysis of the binding sites of autoantibodies have an outstanding importance in understanding the immunopathology of the disease and also in the design of novel diagnostics. Here, we describe the localization of the B-cell epitope regions of Dsg1 and Dsg3 proteins' extracellular parts recognized by IgG-type serum autoantibodies of patients with PV and PF. In our study, overlapping pentadecapeptides were synthesized on hydroxypropyl methacrylate pins based on the results of in silico predictions. To detect the interaction between the serum autoantibodies and the immobilized synthetic peptides, modified Enzyme Linked Immunosorbent Assay (ELISA) was performed with pin-attached peptides testing the serum samples of ten patients and four healthy donors. We identified five possible epitope regions (aa86-110, aa196-220, aa226-250, aa326-340, and aa486-520) within the extracellular part of the Dsg1 and four possible epitope regions (aa64-78, aa330-344, aa375-399, and aa446-460) within that of the Dsg3 protein sequence using these methods. Our data showed that serum autoantibodies of patients, previously identified as Dsg1 and Dsg3 positive, are able to recognize continuous linear epitope regions of both Dsg1 and Dsg3 proteins using pin-bound overlapping peptides in modified ELISAs.
authors.affiliation: Research Group of Peptide Chemistry, Hungarian Academy of Sciences, Eötvös L. University, Budapest 112, PO Box 32 H-1518, Hungary.
authors.forename: Hajnalka
authors.initials: H
authors.lastname: Szabados
authors.forename: Szilvia
authors.initials: S
authors.lastname: Bősze
authors.forename: Pálma
authors.initials: P
authors.lastname: Silló
authors.forename: Sarolta
authors.initials: S
authors.lastname: Kárpáti
authors.forename: Ferenc
authors.initials: F
authors.lastname: Hudecz
authors.forename: Katalin
authors.initials: K
authors.lastname: Uray
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001323:Autoantibodies / Q000097:blood / Q000276:immunology*; D051183:Desmoglein 1 / Q000737:chemistry / Q000276:immunology*; D051184:Desmoglein 3 / Q000737:chemistry / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D005260:Female; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D010392:Pemphigus / Q000097:blood / Q000276:immunology*; D010455:Peptides / Q000737:chemistry / Q000276:immunology*; D055815:Young Adult
chemical_list: D001323:Autoantibodies; C495297:DSG1 protein, human; C071894:DSG3 protein, human; D051183:Desmoglein 1; D051184:Desmoglein 3; D018985:Epitopes, B-Lymphocyte; D055767:Immobilized Proteins; D010455:Peptides