Autoantibodies of primary biliary cirrhosis recognize dihydrolipoamide acetyltransferase and inhibit enzyme function

Judith A Van de Water, D. Fregeau, P. Davis, A. Ansari, D. Danner, Patrick S Leung, R. Coppel, M. Eric Gershwin

Research output: Contribution to journalArticle

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Abstract

Autoantibodies against mitochondria occur in the sera of patients with primary biliary cirrhosis (PBC) with characteristic reactivity to an inner membrane protein of approximately 74 kDa. To precisely define these autoantigens, we recently cloned and sequenced a rat liver cDNA (pRMIT) that encodes for all of the epitopes recognized by Ig to the 74-kDa autoantigen. In the present study we have used this recombinant probe as a tool, in addition to purified enzymes, to demonstrate by immunoblotting that the 74-kDa mitochondrial autoantigen is dihydrolipoamide acetyltransferase (EC 2.3.1.12), the core protein of the pyruvate dehydrogenase complex. Furthermore, and of particular interest, inhibition of pyruvate dehydrogenase enzyme activity was demonstrated after incubation with sera from patients with PBC but not from normal volunteers or patients with chronic active hepatitis. Such inhibition was abrogated by absorption of the PBC sera with an expressing subclone of pRMIT, designated pRMIT-603. Identification of dihydrolipoamide acetyltransferase as the target of autoimmunity in PBC provides a reagent that can be used to determine mechanisms by which this molecule is recognized. It will allow study of whether autoimmune reactivity, at the humoral or T cell level, is the basis for the pathogenesis of PBC. Additionally, such data present evidence of functional inhibition of a critical metabolic enzyme. Dihydrolipoamide acetyltransferase is well-known to mitochondrial biochemistry and, similar to identified autoantigens in other autoimmune diseases, is highly conserved in evolution.

Original languageEnglish (US)
Pages (from-to)2321-2324
Number of pages4
JournalJournal of Immunology
Volume141
Issue number7
StatePublished - 1988

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Dihydrolipoyllysine-Residue Acetyltransferase
Biliary Liver Cirrhosis
Autoantibodies
Autoantigens
Enzymes
Serum
Pyruvate Dehydrogenase Complex
Chronic Hepatitis
Pyruvic Acid
Autoimmunity
Immunoblotting
Biochemistry
Autoimmune Diseases
Epitopes
Healthy Volunteers
Mitochondria
Oxidoreductases
Membrane Proteins
Complementary DNA
T-Lymphocytes

ASJC Scopus subject areas

  • Immunology

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Autoantibodies of primary biliary cirrhosis recognize dihydrolipoamide acetyltransferase and inhibit enzyme function. / Van de Water, Judith A; Fregeau, D.; Davis, P.; Ansari, A.; Danner, D.; Leung, Patrick S; Coppel, R.; Gershwin, M. Eric.

In: Journal of Immunology, Vol. 141, No. 7, 1988, p. 2321-2324.

Research output: Contribution to journalArticle

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abstract = "Autoantibodies against mitochondria occur in the sera of patients with primary biliary cirrhosis (PBC) with characteristic reactivity to an inner membrane protein of approximately 74 kDa. To precisely define these autoantigens, we recently cloned and sequenced a rat liver cDNA (pRMIT) that encodes for all of the epitopes recognized by Ig to the 74-kDa autoantigen. In the present study we have used this recombinant probe as a tool, in addition to purified enzymes, to demonstrate by immunoblotting that the 74-kDa mitochondrial autoantigen is dihydrolipoamide acetyltransferase (EC 2.3.1.12), the core protein of the pyruvate dehydrogenase complex. Furthermore, and of particular interest, inhibition of pyruvate dehydrogenase enzyme activity was demonstrated after incubation with sera from patients with PBC but not from normal volunteers or patients with chronic active hepatitis. Such inhibition was abrogated by absorption of the PBC sera with an expressing subclone of pRMIT, designated pRMIT-603. Identification of dihydrolipoamide acetyltransferase as the target of autoimmunity in PBC provides a reagent that can be used to determine mechanisms by which this molecule is recognized. It will allow study of whether autoimmune reactivity, at the humoral or T cell level, is the basis for the pathogenesis of PBC. Additionally, such data present evidence of functional inhibition of a critical metabolic enzyme. Dihydrolipoamide acetyltransferase is well-known to mitochondrial biochemistry and, similar to identified autoantigens in other autoimmune diseases, is highly conserved in evolution.",
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T1 - Autoantibodies of primary biliary cirrhosis recognize dihydrolipoamide acetyltransferase and inhibit enzyme function

AU - Van de Water, Judith A

AU - Fregeau, D.

AU - Davis, P.

AU - Ansari, A.

AU - Danner, D.

AU - Leung, Patrick S

AU - Coppel, R.

AU - Gershwin, M. Eric

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