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

Hajnalka Szabados, Sz. Bősze, Pálma Silló, S. Kárpáti, F. Hudecz, K. Uray

Research output: Contribution to journalArticle

2 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)84-94
Number of pages11
JournalJournal of Peptide Science
Volume19
Issue number2
DOIs
Publication statusPublished - Feb 2013

Fingerprint

Desmoglein 3
Desmoglein 1
Immobilized Proteins
B-Lymphocyte Epitopes
Pemphigus
Autoantibodies
Peptides
Serum
Epitopes
Proteins
hydroxypropyl methacrylate
Enzyme-Linked Immunosorbent Assay
Immunosorbents
Cell adhesion
Blister
Cell Adhesion
Computer Simulation
Erosion
Assays
Glycoproteins

Keywords

  • Autoantibody epitopes
  • Desmoglein 1 and 3
  • Modified ELISA
  • Pemphigus
  • Pin-bound peptides

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Medicine
  • Molecular Biology
  • Biochemistry
  • Pharmacology
  • Drug Discovery
  • Organic Chemistry

Cite this

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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.",
keywords = "Autoantibody epitopes, Desmoglein 1 and 3, Modified ELISA, Pemphigus, Pin-bound peptides",
author = "Hajnalka Szabados and Sz. Bősze and P{\'a}lma Sill{\'o} and S. K{\'a}rp{\'a}ti and F. Hudecz and K. Uray",
year = "2013",
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T1 - The mapping of linear B-cell epitope regions in the extracellular parts of the desmoglein 1 and 3 proteins

T2 - Recognition of immobilized peptides by pemphigus patients' serum autoantibodies

AU - Szabados, Hajnalka

AU - Bősze, Sz.

AU - Silló, Pálma

AU - Kárpáti, S.

AU - Hudecz, F.

AU - Uray, K.

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N2 - 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.

AB - 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.

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KW - Modified ELISA

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KW - Pin-bound peptides

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