Role of Lipoylation of the Immunodominant Epitope of Pyruvate Dehydrogenase Complex: Toward a Peptide-Based Diagnostic Assay for Primary Biliary Cirrhosis

Giulia Pacini, Alfonso Carotenuto, Cedric Rentier, Francesca Nuti, Feliciana Real-Fernandez, Diego Brancaccio, Giuseppina Sabatino, Maud Larregola, Elisa Peroni, Paola Migliorini, Ettore Novellino, Pier Maria Battezzati, Carlo Selmi, Anna Maria Papini, Paolo Rovero

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Primary biliary cirrhosis is an immune-mediated chronic liver disease whose diagnosis relies on the detection of serum antimitochondrial antibodies directed against a complex set of proteins, among which pyruvate dehydrogenase complex is considered the main autoantigen. We studied the immunological role of the lipoyl domain of this protein using synthetic lipoylated peptides, showing that the lipoyl chain chirality does not affect autoantibody recognition and, most importantly, confirming that both lipoylated and unlipoylated peptides are able to recognize specific autoantibodies in patients sera. In fact, 74% of patients sera recognize at least one of the tested peptides but very few positive sera recognized exclusively the lipoylated peptide, suggesting that the lipoamide moiety plays a marginal role within the autoreactive epitope. These results are supported by a conformational analysis showing that the lipoyl moiety of pyruvate dehydrogenase complex appears to be involved in hydrophobic interactions, which may limit its exposition and thus its contribution to the complex antigenic epitope. A preliminary analysis of the specificity of the two most active peptides indicates that they could be part of a panel of synthetic antigens collectively able to mimic in a simple immunoenzymatic assay the complex positivity pattern detected in immunofluorescence. (Figure Presented).

Original languageEnglish (US)
Pages (from-to)6619-6629
Number of pages11
JournalJournal of Medicinal Chemistry
Volume58
Issue number16
DOIs
StatePublished - Aug 27 2015
Externally publishedYes

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Lipoylation
Pyruvate Dehydrogenase Complex
Immunodominant Epitopes
Biliary Liver Cirrhosis
Peptides
Serum
Autoantibodies
Epitopes
Synthetic Vaccines
Autoantigens
Hydrophobic and Hydrophilic Interactions
Fluorescent Antibody Technique
Liver Diseases
Chronic Disease
Antibodies
Proteins

ASJC Scopus subject areas

  • Molecular Medicine
  • Drug Discovery

Cite this

Role of Lipoylation of the Immunodominant Epitope of Pyruvate Dehydrogenase Complex : Toward a Peptide-Based Diagnostic Assay for Primary Biliary Cirrhosis. / Pacini, Giulia; Carotenuto, Alfonso; Rentier, Cedric; Nuti, Francesca; Real-Fernandez, Feliciana; Brancaccio, Diego; Sabatino, Giuseppina; Larregola, Maud; Peroni, Elisa; Migliorini, Paola; Novellino, Ettore; Battezzati, Pier Maria; Selmi, Carlo; Papini, Anna Maria; Rovero, Paolo.

In: Journal of Medicinal Chemistry, Vol. 58, No. 16, 27.08.2015, p. 6619-6629.

Research output: Contribution to journalArticle

Pacini, G, Carotenuto, A, Rentier, C, Nuti, F, Real-Fernandez, F, Brancaccio, D, Sabatino, G, Larregola, M, Peroni, E, Migliorini, P, Novellino, E, Battezzati, PM, Selmi, C, Papini, AM & Rovero, P 2015, 'Role of Lipoylation of the Immunodominant Epitope of Pyruvate Dehydrogenase Complex: Toward a Peptide-Based Diagnostic Assay for Primary Biliary Cirrhosis', Journal of Medicinal Chemistry, vol. 58, no. 16, pp. 6619-6629. https://doi.org/10.1021/acs.jmedchem.5b00783
Pacini, Giulia ; Carotenuto, Alfonso ; Rentier, Cedric ; Nuti, Francesca ; Real-Fernandez, Feliciana ; Brancaccio, Diego ; Sabatino, Giuseppina ; Larregola, Maud ; Peroni, Elisa ; Migliorini, Paola ; Novellino, Ettore ; Battezzati, Pier Maria ; Selmi, Carlo ; Papini, Anna Maria ; Rovero, Paolo. / Role of Lipoylation of the Immunodominant Epitope of Pyruvate Dehydrogenase Complex : Toward a Peptide-Based Diagnostic Assay for Primary Biliary Cirrhosis. In: Journal of Medicinal Chemistry. 2015 ; Vol. 58, No. 16. pp. 6619-6629.
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abstract = "Primary biliary cirrhosis is an immune-mediated chronic liver disease whose diagnosis relies on the detection of serum antimitochondrial antibodies directed against a complex set of proteins, among which pyruvate dehydrogenase complex is considered the main autoantigen. We studied the immunological role of the lipoyl domain of this protein using synthetic lipoylated peptides, showing that the lipoyl chain chirality does not affect autoantibody recognition and, most importantly, confirming that both lipoylated and unlipoylated peptides are able to recognize specific autoantibodies in patients sera. In fact, 74{\%} of patients sera recognize at least one of the tested peptides but very few positive sera recognized exclusively the lipoylated peptide, suggesting that the lipoamide moiety plays a marginal role within the autoreactive epitope. These results are supported by a conformational analysis showing that the lipoyl moiety of pyruvate dehydrogenase complex appears to be involved in hydrophobic interactions, which may limit its exposition and thus its contribution to the complex antigenic epitope. A preliminary analysis of the specificity of the two most active peptides indicates that they could be part of a panel of synthetic antigens collectively able to mimic in a simple immunoenzymatic assay the complex positivity pattern detected in immunofluorescence. (Figure Presented).",
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AU - Pacini, Giulia

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AU - Rentier, Cedric

AU - Nuti, Francesca

AU - Real-Fernandez, Feliciana

AU - Brancaccio, Diego

AU - Sabatino, Giuseppina

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AU - Peroni, Elisa

AU - Migliorini, Paola

AU - Novellino, Ettore

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AU - Selmi, Carlo

AU - Papini, Anna Maria

AU - Rovero, Paolo

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N2 - Primary biliary cirrhosis is an immune-mediated chronic liver disease whose diagnosis relies on the detection of serum antimitochondrial antibodies directed against a complex set of proteins, among which pyruvate dehydrogenase complex is considered the main autoantigen. We studied the immunological role of the lipoyl domain of this protein using synthetic lipoylated peptides, showing that the lipoyl chain chirality does not affect autoantibody recognition and, most importantly, confirming that both lipoylated and unlipoylated peptides are able to recognize specific autoantibodies in patients sera. In fact, 74% of patients sera recognize at least one of the tested peptides but very few positive sera recognized exclusively the lipoylated peptide, suggesting that the lipoamide moiety plays a marginal role within the autoreactive epitope. These results are supported by a conformational analysis showing that the lipoyl moiety of pyruvate dehydrogenase complex appears to be involved in hydrophobic interactions, which may limit its exposition and thus its contribution to the complex antigenic epitope. A preliminary analysis of the specificity of the two most active peptides indicates that they could be part of a panel of synthetic antigens collectively able to mimic in a simple immunoenzymatic assay the complex positivity pattern detected in immunofluorescence. (Figure Presented).

AB - Primary biliary cirrhosis is an immune-mediated chronic liver disease whose diagnosis relies on the detection of serum antimitochondrial antibodies directed against a complex set of proteins, among which pyruvate dehydrogenase complex is considered the main autoantigen. We studied the immunological role of the lipoyl domain of this protein using synthetic lipoylated peptides, showing that the lipoyl chain chirality does not affect autoantibody recognition and, most importantly, confirming that both lipoylated and unlipoylated peptides are able to recognize specific autoantibodies in patients sera. In fact, 74% of patients sera recognize at least one of the tested peptides but very few positive sera recognized exclusively the lipoylated peptide, suggesting that the lipoamide moiety plays a marginal role within the autoreactive epitope. These results are supported by a conformational analysis showing that the lipoyl moiety of pyruvate dehydrogenase complex appears to be involved in hydrophobic interactions, which may limit its exposition and thus its contribution to the complex antigenic epitope. A preliminary analysis of the specificity of the two most active peptides indicates that they could be part of a panel of synthetic antigens collectively able to mimic in a simple immunoenzymatic assay the complex positivity pattern detected in immunofluorescence. (Figure Presented).

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