The mapping of linear B-cell epitope regions in desmoglein 1 and 3 proteins : Recognition of immobilized peptides by pemphigus patients’ serum autoantibodies

Szabados, Hajnalka and Bősze, Szilvia and Silló, Pálma and Kárpáti, Sarolta and Hudecz, Ferenc and Uray, Katalin (2013) The mapping of linear B-cell epitope regions in desmoglein 1 and 3 proteins : Recognition of immobilized peptides by pemphigus patients’ serum autoantibodies. JOURNAL OF PEPTIDE SCIENCE, 19 (2). pp. 84-94. ISSN 1075-2617

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Abstract

Desmosomal transmembrane glycoproteins desmoglein 1and desmoglein 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 theimmunopathology 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 andPF. In our study overlapping pentadecapeptides were synthesized on hydroxypropylmethacrylate pins based on the results of in silicopredictions. To detect the interaction between theserum autoantibodies and the immobilized synthetic peptides, modified ELISA (Enzyme Linked Immunosorbent Assay) 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 Dsg1 protein sequence and four possible epitope regions (aa64-78, aa330-344, aa375-399, aa446-460) within 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.

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