Conformational epitopes of pemphigus antigens (Dsg1 and dsg3) are calcium dependent and glycosylation independent

Masayuki Amagai, K. Ishii, T. Hashimoto, S. Gamou, N. Shimizu, T. Nishikawa

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

The target molecule of pemphigus autoantibodies is a transmembrane desmosomal component, desmoglein 3 (Dsg3) in pemphigus vulgaris (PV) and Dsg1 in pemphigus foliaceus (PF). In this study, we examined the effects of calcium and glycosylation on the antigenicity of the pemphigus antigens and on the generation of conformational epitopes. We used recombinant baculovirus proteins, PVIg and PFIg, which are considered to reflect accurately the native conformation of the extracellular domain of their respective proteins Dsg3 and Dsg1. These baculoproteins could immunoadsorb heterogeneous autoantibodies from the corresponding sera of PV and PF patients, completely blocking indirect immunofluorescence staining of normal human skin. Chelating calcium from the solution containing the baculoproteins using ethylenediaminetetraacetic acid (EDTA) or ethyleneglycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) abolished immunoadsorption by both PVIg and PFIg; however, immunoadsorption by the baculoproteins was restored after dialysis against 1 mM calcium. Nonglycosylated forms of both baculoproteins produced in the presence of tunicamycin retained their immunoadsorptive ability. Furthermore, immunoadsorption by the baculoproteins was prevented irreversibly by treatment with low pH, high pH, and boiling, but not with the non-ionic detergent Nonidet P-40. These findings indicate that formation of the conformational epitopes on the pemphigus antigens is dependent on calcium but independent of glycosylation, and provide direct evidence that calcium plays an important role in determining the antigenic properties of the pemphigus antigens.

Original languageEnglish
Pages (from-to)243-247
Number of pages5
JournalJournal of Investigative Dermatology
Volume105
Issue number2
Publication statusPublished - 1995
Externally publishedYes

Fingerprint

Desmoglein 1
Glycosylation
Pemphigus
Epitopes
Calcium
Desmoglein 3
Antigens
Autoantibodies
Tunicamycin
Recombinant proteins
Dialysis
Egtazic Acid
Chelation
Edetic Acid
Detergents
Ether
Boiling liquids
Conformations
Skin
Baculoviridae

Keywords

  • Autoimmune disease
  • Baculovirus expression
  • Cadherin
  • Desmoglein

ASJC Scopus subject areas

  • Dermatology

Cite this

Conformational epitopes of pemphigus antigens (Dsg1 and dsg3) are calcium dependent and glycosylation independent. / Amagai, Masayuki; Ishii, K.; Hashimoto, T.; Gamou, S.; Shimizu, N.; Nishikawa, T.

In: Journal of Investigative Dermatology, Vol. 105, No. 2, 1995, p. 243-247.

Research output: Contribution to journalArticle

Amagai, Masayuki ; Ishii, K. ; Hashimoto, T. ; Gamou, S. ; Shimizu, N. ; Nishikawa, T. / Conformational epitopes of pemphigus antigens (Dsg1 and dsg3) are calcium dependent and glycosylation independent. In: Journal of Investigative Dermatology. 1995 ; Vol. 105, No. 2. pp. 243-247.
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