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eDNA Cloning of the 210-kDa Paraneoplastic Pemphigus Antigen Reveals that Envoplakin Is a Component of the Antigen Complex

Authors
 Soo-Chan Kim  ;  Young Do Kwon  ;  In Joon Lee IlJoo Lee  ;  Sung Nam Chang  ;  Tae Gyu Lee 
Citation
 JOURNAL OF INVESTIGATIVE DERMATOLOGY, Vol.109(3) : 365-369, 1997 
Journal Title
 JOURNAL OF INVESTIGATIVE DERMATOLOGY 
ISSN
 0022-202X 
Issue Date
1997
MeSH
Autoantibodies/analysis ; Autoantigens/chemistry ; Base Sequence ; Cadherins/chemistry* ; Cloning, Molecular ; DNA, Complementary/analysis ; Desmoglein 1 ; Desmoglein 3 ; Gene Library ; Humans ; Membrane Proteins/analysis* ; Molecular Sequence Data ; Paraneoplastic Syndromes/immunology ; Pemphigus/immunology ; Protein Precursors/analysis* ; Sequence Analysis, DNA
Abstract
Although the 210 and 190-kDa proteins are the most frequently detected antigens reacting with sera of patients with paraneoplastic pemphigus (PNP) in immunoblot analysis, there is still uncertainty as to the nature of these PNP antigens. To isolate and characterize a cDNA clone encoding the 210-kDa PNP antigen, we screened a human keratinocyte lambda gt 11 cDNA expression library by the immunoperoxidase method with serum IgG from a PNP patient. The IgG used for the immunoscreening of a keratinocyte cDNA expression library recognized 210- and 190-kDa antigens by immunoblotting. A single clone, called here the PNP clone, producing a fusion protein that reacted strongly with the patient's IgG, was further characterized. Only the PNP patient's IgG, but not IgG from a normal control, pemphigus foliaceus, or pemphigus vulgaris patients, bound the plaques of this positive clone. Furthermore, PNP IgG affinity purified on plaques of this clone, but not unrelated clones, bound to keratinocyte cell surfaces by immunofluorescence and reacted with the 210-kDa PNP antigen by immunoblotting. EcoRI digestion of the clone's cDNA insert demonstrated a 1.4-kbp fragment. This cDNA insert was placed into a M13 mp 18 vector and sequenced. Sequence analysis revealed that the cDNA insert of the PNP clone encodes a part of the central rod domain and the COOH-terminal C domain of envoplakin, a newly defined precursor of the cornified envelope that is homologous to desmoplakin. This result demonstrates that the 210-kDa PNP antigen is envoplakin and PNP is an autoimmune disease that produces autoantibodies against intermediate filament-associated proteins in desmosomes and hemidesmosomes, desmoplakin, bullous pemphigoid antigen 1 (BPAG 1), and envoplakin.
Files in This Item:
T199701899.pdf Download
DOI
10.1111/1523-1747.ep12336235
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Dermatology (피부과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Soo Chan(김수찬) ORCID logo https://orcid.org/0000-0002-2327-4755
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/177327
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