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Epitope mapping of mitochondrial adenine nucleotide translocase-1 in idiopathic dilated cardiomyopathy.

Abstract
Mitochondrial adenine nucleotide translocase (ANT) is a specific target for the autoantibody response in idiopathic dilated cardiomyopathy (IDCM). We have undertaken an epitope analysis of ANT in IDCM by immunoblot with recombinant GST-ANT fusion proteins and with cellulose-bound decapeptides of human ANT1. Forty-five patients with IDCM, 17 patients with ischemic left ventricle dysfunction (LVD) and 20 controls were analyzed for circulating antibodies against ANT (AAb-ANT). Sixteen of the 45 (36%) IDCM patients showed AAb-ANT above controls. In immunoblots, AAb-ANT detected purified bovine heart ANT and GST-ANT1 and GST-ANT2 isoforms and, less frequently, the GST-ANT3 isoform. A construct lacking the last 146 amino acids did not react with AAb-ANT, indicating that the main epitopes are in the C-terminal 146 amino acids. Immunodetection of decapeptides covering this region shows that AAb-ANT detects at least three epitopes, demonstrating that ANT is the primary target of AAb-ANT. The most significant epitopes belong to the M2 and M3 hydrophilic loops of ANT suggesting that apart from being essential for its activity, these loops are highly immunogenic.
AuthorsCarlos Manchado, Josefina Orús, Francesc Villarroya, Eulalia Roig, Magda Heras, Marta Giralt, Roser Iglesias, Ginés Sanz, Teresa Mampel, Octavi Viñas
JournalJournal of molecular and cellular cardiology (J Mol Cell Cardiol) Vol. 34 Issue 5 Pg. 571-82 (May 2002) ISSN: 0022-2828 [Print] England
PMID12056860 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright 2002 Academic Press.
Chemical References
  • Autoantigens
  • Protein Isoforms
  • Recombinant Fusion Proteins
  • Mitochondrial ADP, ATP Translocases
Topics
  • Animals
  • Autoantigens (immunology)
  • Cardiomyopathy, Dilated (enzymology, immunology)
  • Cattle
  • Cloning, Molecular
  • Enzyme-Linked Immunosorbent Assay
  • Epitope Mapping
  • Female
  • Humans
  • Immunoblotting
  • Male
  • Middle Aged
  • Mitochondria (immunology, metabolism)
  • Mitochondrial ADP, ATP Translocases (immunology)
  • Protein Isoforms
  • Rats
  • Recombinant Fusion Proteins

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