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Characterization of a Novel Isoform of α-Nascent Polypeptide-associated Complex as IgE-defined Autoantigen
Journal article   Open access   Peer reviewed

Characterization of a Novel Isoform of α-Nascent Polypeptide-associated Complex as IgE-defined Autoantigen

Roschanak Mossabeb, Susanne Seiberler, Irene Mittermann, Susanne Natter, Dietrich Kraft, Rudolf Valenta, Renate Reininger, Susanne Spitzauer, Petra Verdino and Walter Keller
Journal of investigative dermatology, v 119(4), pp 820-829
01 Oct 2002
PMID: 12406326
url
https://doi.org/10.1046/j.1523-1747.2002.00518.xView
Published, Version of Record (VoR) Open

Abstract

allergy atopic dermatitis autoantigen circular dichroism nascent polypeptide-associated complex
The nascent polypeptide-associated complex is required for intracellular translocation of newly synthesized polypeptides in eukaryotic cells. It may also act as a transcriptional coactivator in humans and various eukaryotic organisms and binds to nucleic acids. Recently, we provided evidence that a component of nascent polypeptide-associated complex, α-nascent polypeptide-associated complex, represents an IgE-reactive autoantigen for atopic dermatitis patients. By oligonucleotide screening we isolated a complete cDNA coding for a so far unknown α-nascent polypeptide-associated complex isoform from a human epithelial cDNA library. Southern blot hybridization experiments provided further evidence that α-nascent polypeptide-associated complex is encoded by a gene family. Recombinant α-nascent polypeptide-associated complex was expressed in Escherichia coli as a soluble, His-tagged protein, and purified via nickel affinity chromatography. By circular dichroism analysis it is demonstrated that purified recombinant α-nascent polypeptide-associated complex represents a folded protein of mixed α-helical and β-sheet conformation with unusual high thermal stability and remarkable refolding capacity. Complete recombinant α-nascent polypeptide-associated complex (215 amino acids) and its 86 amino acid C-terminal fragment specifically bound IgE autoantibodies. Recombinant α-nascent polypeptide-associated complex also inhibited IgE binding to natural α-nascent polypeptide-associated complex, demonstrating the presence of common IgE epitopes between the recombinant and natural protein. Furthermore, recombinant α-nascent polypeptide-associated complex induced specific lymphoproliferative responses in peripheral blood mononuclear cells of a sensitized atopic dermatitis patient. As has been proposed for environmental allergens it is possible that T cell responses to IgE-defined autoantigens may contribute to the chronic skin manifestations in atopic dermatitis.

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Collaboration types
Domestic collaboration
Web of Science research areas
Dermatology
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