Structural requirement for autoreactivity on human pyruvate dehydrogenase-e2, the major autoantigen of primary biliary cirrhosis implication for a conformational autoepitope

Charles D. Surh, Ross Coppel, M. Eric Gershwin

Research output: Contribution to journalArticle

145 Scopus citations

Abstract

The E2 component (acetyltransferase) of the pyruvate dehydrogenase (PDH) complex is the major mitochondrial autoantigen recognized by autoantibodies in patients with primary biliary cirrhosis (PBC). Previous work, using only a partial length rat liver cDNA clone of PDH-E2, demonstrated that the immunodominant epitope was localized to the lipoic acid binding site. Human PDH-E2, in contrast to rat PDH-E2, has two lipoic acid binding sites. By using a full length human cDNA for PDH-E2, and by preparation of multiple overlapping recombinant fragments, we have determined that three autoreactive determinants are present on human PDH-E2: two cross-reactive lipoyl domains, and an area surrounding the E1/E3 binding region. The dominant epitope was localized to the inner lipoyl domain whereas the outer lipoyl domain only showed a weak cross-reactivity, and only 1/26 PBC sera reacted weakly to the E1/E3 binding region area. By probing recombinant fusion proteins expressed from small restriction fragments of the inner lipoyl domain, we have found that a minimum of 75 amino acids (residues 146-221) were required for detectable autoantibody binding, and that 93 amino acids (residues 128-221) were necessary for characteristically strong antimitochondrial autoantibody recognition. Such a requirement for a large region suggests the possibility that a conformational autoepitope may be recognized. In addition, we have found that absorption of PBC sera with the purified mammalian PDH complex does not remove reactivity against Escherichia coli Ag. The possible implications for such results are discussed.

Original languageEnglish (US)
Pages (from-to)3367-3374
Number of pages8
JournalJournal of Immunology
Volume144
Issue number9
StatePublished - May 1 1990

ASJC Scopus subject areas

  • Immunology

Fingerprint Dive into the research topics of 'Structural requirement for autoreactivity on human pyruvate dehydrogenase-e2, the major autoantigen of primary biliary cirrhosis implication for a conformational autoepitope'. Together they form a unique fingerprint.

  • Cite this