{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T18:53:23Z","timestamp":1777575203174,"version":"3.51.4"},"reference-count":39,"publisher":"Oxford University Press (OUP)","issue":"5","license":[{"start":{"date-parts":[[2013,9,1]],"date-time":"2013-09-01T00:00:00Z","timestamp":1377993600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2013,9,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Antimitochondrial autoantibodies (AMAs), the serological hallmark of primary biliary cirrhosis, are directed against the lipoyl domain of the E2 subunit of pyruvate dehydrogenase (PDC-E2). However, comprehensive analysis of the amino acid residues of PDC-E2 lipoyl \u03b2-sheet with AMA specificity is lacking. In this study, we postulated that specific residues within the lipoyl domain are critical to AMA recognition by maintaining conformational integrity. We systematically replaced each of 19 residue peptides of the inner lipoyl domain with alanine and analyzed these mutants for reactivities against 60 primary biliary cirrhosis and 103 control sera. Based on these data, we then constructed mutants with two, three, or four replacements and, in addition, probed the structure of the substituted domains using thiol-specific spin labeling and electron paramagnetic resonance (EPR) of a 5Ile\u2192Ala and 12Ile\u2192Ala double mutant. Single alanine replacement at 5Ile, 12Ile, and 15Glu significantly reduced AMA recognition. In addition, mutants with two, three, or four replacements at 5Ile, 12Ile, and 15Glu reduced AMA reactivity even further. Indeed, EPR reveals a highly flexible structure within the 5Ile and 12Ile double-alanine mutant. Autoreactivity is largely focused on specific residues in the PDC-E2 lipoyl domain critical in maintaining the lipoyl loop conformation necessary for AMA recognition. Collectively, the AMA binding studies and EPR analysis demonstrate the necessity of the lipoyl \u03b2-sheet structural conformation in anti\u2013PDC-E2 recognition.<\/jats:p>","DOI":"10.4049\/jimmunol.1301092","type":"journal-article","created":{"date-parts":[[2013,7,27]],"date-time":"2013-07-27T06:41:57Z","timestamp":1374907317000},"page":"2126-2133","source":"Crossref","is-referenced-by-count":31,"title":["Antimitochondrial Antibody Recognition and Structural Integrity of the Inner Lipoyl Domain of the E2 Subunit of Pyruvate Dehydrogenase Complex"],"prefix":"10.1093","volume":"191","author":[{"given":"Jinjun","family":"Wang","sequence":"first","affiliation":[{"name":"Division of Rheumatology, Allergy and Clinical Immunology, University of California Davis , Davis, CA 95616"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Madhu S","family":"Budamagunta","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Medicine, School of Medicine, University of California Davis , Davis, CA 95616"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John C","family":"Voss","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Medicine, School of Medicine, University of California Davis , Davis, CA 95616"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark J","family":"Kurth","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of California Davis , Davis, CA 95616"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kit S","family":"Lam","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Medicine, University of California Davis , Davis, CA 95817"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ling","family":"Lu","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Nanjing Normal University , Nanjing 210023,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas P","family":"Kenny","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Allergy and Clinical Immunology, University of California Davis , Davis, CA 95616"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher","family":"Bowlus","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology and Hepatology, University of California Davis Medical Center , Sacramento, CA 95817"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kentaro","family":"Kikuchi","sequence":"additional","affiliation":[{"name":"The Fourth Department of Internal Medicine, Teikyo University , Kanagawa 213-8507,","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ross L","family":"Coppel","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Monash University , Clayton, Victoria 3800,","place":["Australia"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aftab A","family":"Ansari","sequence":"additional","affiliation":[{"name":"Department of Pathology, Emory University School of Medicine , Atlanta, GA 30322"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M Eric","family":"Gershwin","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Allergy and Clinical Immunology, University of California Davis , Davis, CA 95616"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patrick S C","family":"Leung","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Allergy and Clinical Immunology, University of California Davis , Davis, CA 95616"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2013,9,1]]},"reference":[{"key":"2025032405274187500_r1","doi-asserted-by":"crossref","first-page":"737","DOI":"10.1002\/hep.22042","article-title":"The causes of primary biliary cirrhosis: convenient and inconvenient truths","volume":"47","author":"Gershwin","year":"2008","journal-title":"Hepatology"},{"key":"2025032405274187500_r2","doi-asserted-by":"crossref","first-page":"3525","DOI":"10.4049\/jimmunol.138.10.3525","article-title":"Identification and specificity of a cDNA encoding the 70 kd mitochondrial antigen recognized in primary biliary cirrhosis","volume":"138","author":"Gershwin","year":"1987","journal-title":"J. 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