Epitope mapping and reactivity of autoantibodies to the E2 component of 2-oxoglutarate dehydrogenase complex in primary biliary cirrhosis using recombinant 2-oxoglutarate dehydrogenase complex

Sekio Moteki, Patrick S Leung, E. Rolland Dickson, David H. Van Thiel, Claudio Galperin, Thorsten Buch, Donato Alarcon-Segovia, David Kershenobich, Kinya Kawano, Ross L. Coppel, Sadayuki Matuda, M. Eric Gershwin

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Abstract

Five different target mitochondrial autoantigens recognized by sera from patients with primary biliary cirrhosis (PBC) have been identified as subunits of the following 2-oxo acid dehydrogenase complexes: the pyruvate dehydrogenase complex (PDC), the branched chain 2-oxo acid dehydrogenase complex (BCOADC), and the 2-oxoglutarate dehydrogenase complex (OGDC). Unlike the E2 subunits of PDC (PDC-E2) and BCOADC (BCOADC-E2), the E2 subunits of OGDC (OGDC-E2) reactivity of PBC sera and the reactive epitope of OGDC-D2 have not hitherto been studied in detail. In this report, we took advantage of a recombinant fusion protein for OGDC-E2 to address these issues. Eighty of 268 (29.9%) PBC patient sera but none of 45 controls reacted with recombinant OGDC-E2. The recombinant OGDC-E2 was judged to express the immunodominant epitope, because when sera from patients with PBC were preabsorbed with the recombinant fusion protein, such sera were depleted of reactivity against 48 kD OGDC-E2 when probed on beef heart mitochondria (BHM) but retained reactivity toward PDC-E2 and/or BCOADC-E2. Furthermore, affinity-purified PBC sera against recombinant OGDC-E2 reacted only with native OGDC-E2 and not with any other enzyme components of the 2-oxo acid dehydrogenase complex. Antimitochondrial autoantibodies (AMA) against OGDC-E2 included immunoglobulin (Ig) G2, IgG3 and IgM and the relative titers were as follows: IgG2 >IgG3 >IgM. Finally, using overlapping recombinant polypeptides, it was determined that a minimum of 81 amino acids (residues 67-147) corresponding to the lipoyl domain of OGDC-E2 are necessary for reactivity, suggesting that a conformational autoepitope is recognized by AMA. These data suggest that each of the 2-oxo acid dehydrogenase enzymes has distinct antigenicity despite their similarities in structure and function. The availability of recombinant OGDC-E2 autoantigen will allow the design of additional studies to further our understanding of the role of mitochondrial autoantigens in the pathogenesis of PBC.

Original languageEnglish (US)
Pages (from-to)436-444
Number of pages9
JournalHepatology
Volume23
Issue number3
StatePublished - 1996

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Ketoglutarate Dehydrogenase Complex
Epitope Mapping
Biliary Liver Cirrhosis
Autoantibodies
3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)
Dihydrolipoyllysine-Residue Acetyltransferase
Keto Acids
Autoantigens
Serum
Recombinant Fusion Proteins
Oxidoreductases
Immunoglobulin G
Immunoglobulin M
Pyruvate Dehydrogenase Complex
Heart Mitochondria
Immunodominant Epitopes
Enzymes

ASJC Scopus subject areas

  • Hepatology

Cite this

Epitope mapping and reactivity of autoantibodies to the E2 component of 2-oxoglutarate dehydrogenase complex in primary biliary cirrhosis using recombinant 2-oxoglutarate dehydrogenase complex. / Moteki, Sekio; Leung, Patrick S; Rolland Dickson, E.; Van Thiel, David H.; Galperin, Claudio; Buch, Thorsten; Alarcon-Segovia, Donato; Kershenobich, David; Kawano, Kinya; Coppel, Ross L.; Matuda, Sadayuki; Gershwin, M. Eric.

In: Hepatology, Vol. 23, No. 3, 1996, p. 436-444.

Research output: Contribution to journalArticle

Moteki, S, Leung, PS, Rolland Dickson, E, Van Thiel, DH, Galperin, C, Buch, T, Alarcon-Segovia, D, Kershenobich, D, Kawano, K, Coppel, RL, Matuda, S & Gershwin, ME 1996, 'Epitope mapping and reactivity of autoantibodies to the E2 component of 2-oxoglutarate dehydrogenase complex in primary biliary cirrhosis using recombinant 2-oxoglutarate dehydrogenase complex', Hepatology, vol. 23, no. 3, pp. 436-444.
Moteki, Sekio ; Leung, Patrick S ; Rolland Dickson, E. ; Van Thiel, David H. ; Galperin, Claudio ; Buch, Thorsten ; Alarcon-Segovia, Donato ; Kershenobich, David ; Kawano, Kinya ; Coppel, Ross L. ; Matuda, Sadayuki ; Gershwin, M. Eric. / Epitope mapping and reactivity of autoantibodies to the E2 component of 2-oxoglutarate dehydrogenase complex in primary biliary cirrhosis using recombinant 2-oxoglutarate dehydrogenase complex. In: Hepatology. 1996 ; Vol. 23, No. 3. pp. 436-444.
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abstract = "Five different target mitochondrial autoantigens recognized by sera from patients with primary biliary cirrhosis (PBC) have been identified as subunits of the following 2-oxo acid dehydrogenase complexes: the pyruvate dehydrogenase complex (PDC), the branched chain 2-oxo acid dehydrogenase complex (BCOADC), and the 2-oxoglutarate dehydrogenase complex (OGDC). Unlike the E2 subunits of PDC (PDC-E2) and BCOADC (BCOADC-E2), the E2 subunits of OGDC (OGDC-E2) reactivity of PBC sera and the reactive epitope of OGDC-D2 have not hitherto been studied in detail. In this report, we took advantage of a recombinant fusion protein for OGDC-E2 to address these issues. Eighty of 268 (29.9{\%}) PBC patient sera but none of 45 controls reacted with recombinant OGDC-E2. The recombinant OGDC-E2 was judged to express the immunodominant epitope, because when sera from patients with PBC were preabsorbed with the recombinant fusion protein, such sera were depleted of reactivity against 48 kD OGDC-E2 when probed on beef heart mitochondria (BHM) but retained reactivity toward PDC-E2 and/or BCOADC-E2. Furthermore, affinity-purified PBC sera against recombinant OGDC-E2 reacted only with native OGDC-E2 and not with any other enzyme components of the 2-oxo acid dehydrogenase complex. Antimitochondrial autoantibodies (AMA) against OGDC-E2 included immunoglobulin (Ig) G2, IgG3 and IgM and the relative titers were as follows: IgG2 >IgG3 >IgM. Finally, using overlapping recombinant polypeptides, it was determined that a minimum of 81 amino acids (residues 67-147) corresponding to the lipoyl domain of OGDC-E2 are necessary for reactivity, suggesting that a conformational autoepitope is recognized by AMA. These data suggest that each of the 2-oxo acid dehydrogenase enzymes has distinct antigenicity despite their similarities in structure and function. The availability of recombinant OGDC-E2 autoantigen will allow the design of additional studies to further our understanding of the role of mitochondrial autoantigens in the pathogenesis of PBC.",
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T1 - Epitope mapping and reactivity of autoantibodies to the E2 component of 2-oxoglutarate dehydrogenase complex in primary biliary cirrhosis using recombinant 2-oxoglutarate dehydrogenase complex

AU - Moteki, Sekio

AU - Leung, Patrick S

AU - Rolland Dickson, E.

AU - Van Thiel, David H.

AU - Galperin, Claudio

AU - Buch, Thorsten

AU - Alarcon-Segovia, Donato

AU - Kershenobich, David

AU - Kawano, Kinya

AU - Coppel, Ross L.

AU - Matuda, Sadayuki

AU - Gershwin, M. Eric

PY - 1996

Y1 - 1996

N2 - Five different target mitochondrial autoantigens recognized by sera from patients with primary biliary cirrhosis (PBC) have been identified as subunits of the following 2-oxo acid dehydrogenase complexes: the pyruvate dehydrogenase complex (PDC), the branched chain 2-oxo acid dehydrogenase complex (BCOADC), and the 2-oxoglutarate dehydrogenase complex (OGDC). Unlike the E2 subunits of PDC (PDC-E2) and BCOADC (BCOADC-E2), the E2 subunits of OGDC (OGDC-E2) reactivity of PBC sera and the reactive epitope of OGDC-D2 have not hitherto been studied in detail. In this report, we took advantage of a recombinant fusion protein for OGDC-E2 to address these issues. Eighty of 268 (29.9%) PBC patient sera but none of 45 controls reacted with recombinant OGDC-E2. The recombinant OGDC-E2 was judged to express the immunodominant epitope, because when sera from patients with PBC were preabsorbed with the recombinant fusion protein, such sera were depleted of reactivity against 48 kD OGDC-E2 when probed on beef heart mitochondria (BHM) but retained reactivity toward PDC-E2 and/or BCOADC-E2. Furthermore, affinity-purified PBC sera against recombinant OGDC-E2 reacted only with native OGDC-E2 and not with any other enzyme components of the 2-oxo acid dehydrogenase complex. Antimitochondrial autoantibodies (AMA) against OGDC-E2 included immunoglobulin (Ig) G2, IgG3 and IgM and the relative titers were as follows: IgG2 >IgG3 >IgM. Finally, using overlapping recombinant polypeptides, it was determined that a minimum of 81 amino acids (residues 67-147) corresponding to the lipoyl domain of OGDC-E2 are necessary for reactivity, suggesting that a conformational autoepitope is recognized by AMA. These data suggest that each of the 2-oxo acid dehydrogenase enzymes has distinct antigenicity despite their similarities in structure and function. The availability of recombinant OGDC-E2 autoantigen will allow the design of additional studies to further our understanding of the role of mitochondrial autoantigens in the pathogenesis of PBC.

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