Mapping of citrullinated fibrinogen B-cell epitopes in rheumatoid arthritis by imaging surface plasmon resonance

Joyce J.B.C. van Beers, Reinout Raijmakers, Lou-Ella Alexander, Judith Stammen-Vogelzangs, Angelique M.C. Lokate, Albert J.R. Heck, Richardus B.M. Schasfoort, Ger J.M. Pruijn

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Abstract

Introduction Rheumatoid arthritis (RA) frequently involves the loss of tolerance to citrullinated antigens, which may play a role in pathogenicity. Citrullinated fibrinogen is commonly found in inflamed synovial tissue and is a frequent target of autoantibodies in RA patients. To obtain insight into the B-cell response to citrullinated fibrinogen in RA, its autoepitopes were systematically mapped using a new methodology. Methods Human fibrinogen was citrullinated in vitro by peptidylarginine deiminases (PAD), subjected to proteolysis and the resulting peptides were fractionated by ion exchange chromatography. The peptide composition of the citrullinated peptide-containing fractions was determined by high resolution tandem mass spectrometry. The recognition of these fractions by patient sera was subsequently analyzed by imaging surface plasmon resonance on microarrays. Results In total about two-thirds of the 81 arginines of human fibrinogen were found to be susceptible to citrullination by the human PAD2, the human PAD4 or the rabbit PAD2 enzymes. Citrullination sites were found in all three polypeptide chains of fibrinogen, although the α-chain appeared to contain most of them. The analysis of 98 anti-citrullinated protein antibody-positive RA sera using the new methodology allowed the identification of three major citrullinated epitope regions in human fibrinogen, two in the α- and one in the β-chain. Conclusions A comprehensive overview of citrullination sites in human fibrinogen was generated. The multiplex analysis of peptide fractions derived from a post-translationally modified protein, characterized by mass spectrometry, with patient sera provides a versatile system for mapping modified amino acid-containing epitopes. The citrullinated epitopes of human fibrinogen most efficiently recognized by RA autoantibodies are confined to three regions of its polypeptides.
Original languageEnglish
Article numberR219
Pages (from-to)-
Number of pages10
JournalArthritis research & therapy
Volume12
Issue number6
DOIs
Publication statusPublished - 2010

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B-Lymphocyte Epitopes
Surface Plasmon Resonance
Fibrinogen
Rheumatoid Arthritis
Peptides
Epitopes
Autoantibodies
Serum
Ion Exchange Chromatography
Tandem Mass Spectrometry
Proteolysis
Arthritis
Virulence
Arginine
Mass Spectrometry
Proteins
B-Lymphocytes
Rabbits
Antigens
Amino Acids

Keywords

  • METIS-282487
  • IR-104395

Cite this

van Beers, J. J. B. C., Raijmakers, R., Alexander, L-E., Stammen-Vogelzangs, J., Lokate, A. M. C., Heck, A. J. R., ... Pruijn, G. J. M. (2010). Mapping of citrullinated fibrinogen B-cell epitopes in rheumatoid arthritis by imaging surface plasmon resonance. Arthritis research & therapy, 12(6), -. [R219]. https://doi.org/10.1186/ar3205
van Beers, Joyce J.B.C. ; Raijmakers, Reinout ; Alexander, Lou-Ella ; Stammen-Vogelzangs, Judith ; Lokate, Angelique M.C. ; Heck, Albert J.R. ; Schasfoort, Richardus B.M. ; Pruijn, Ger J.M. / Mapping of citrullinated fibrinogen B-cell epitopes in rheumatoid arthritis by imaging surface plasmon resonance. In: Arthritis research & therapy. 2010 ; Vol. 12, No. 6. pp. -.
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title = "Mapping of citrullinated fibrinogen B-cell epitopes in rheumatoid arthritis by imaging surface plasmon resonance",
abstract = "Introduction Rheumatoid arthritis (RA) frequently involves the loss of tolerance to citrullinated antigens, which may play a role in pathogenicity. Citrullinated fibrinogen is commonly found in inflamed synovial tissue and is a frequent target of autoantibodies in RA patients. To obtain insight into the B-cell response to citrullinated fibrinogen in RA, its autoepitopes were systematically mapped using a new methodology. Methods Human fibrinogen was citrullinated in vitro by peptidylarginine deiminases (PAD), subjected to proteolysis and the resulting peptides were fractionated by ion exchange chromatography. The peptide composition of the citrullinated peptide-containing fractions was determined by high resolution tandem mass spectrometry. The recognition of these fractions by patient sera was subsequently analyzed by imaging surface plasmon resonance on microarrays. Results In total about two-thirds of the 81 arginines of human fibrinogen were found to be susceptible to citrullination by the human PAD2, the human PAD4 or the rabbit PAD2 enzymes. Citrullination sites were found in all three polypeptide chains of fibrinogen, although the α-chain appeared to contain most of them. The analysis of 98 anti-citrullinated protein antibody-positive RA sera using the new methodology allowed the identification of three major citrullinated epitope regions in human fibrinogen, two in the α- and one in the β-chain. Conclusions A comprehensive overview of citrullination sites in human fibrinogen was generated. The multiplex analysis of peptide fractions derived from a post-translationally modified protein, characterized by mass spectrometry, with patient sera provides a versatile system for mapping modified amino acid-containing epitopes. The citrullinated epitopes of human fibrinogen most efficiently recognized by RA autoantibodies are confined to three regions of its polypeptides.",
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van Beers, JJBC, Raijmakers, R, Alexander, L-E, Stammen-Vogelzangs, J, Lokate, AMC, Heck, AJR, Schasfoort, RBM & Pruijn, GJM 2010, 'Mapping of citrullinated fibrinogen B-cell epitopes in rheumatoid arthritis by imaging surface plasmon resonance' Arthritis research & therapy, vol. 12, no. 6, R219, pp. -. https://doi.org/10.1186/ar3205

Mapping of citrullinated fibrinogen B-cell epitopes in rheumatoid arthritis by imaging surface plasmon resonance. / van Beers, Joyce J.B.C.; Raijmakers, Reinout; Alexander, Lou-Ella; Stammen-Vogelzangs, Judith; Lokate, Angelique M.C.; Heck, Albert J.R.; Schasfoort, Richardus B.M.; Pruijn, Ger J.M.

In: Arthritis research & therapy, Vol. 12, No. 6, R219, 2010, p. -.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Mapping of citrullinated fibrinogen B-cell epitopes in rheumatoid arthritis by imaging surface plasmon resonance

AU - van Beers, Joyce J.B.C.

AU - Raijmakers, Reinout

AU - Alexander, Lou-Ella

AU - Stammen-Vogelzangs, Judith

AU - Lokate, Angelique M.C.

AU - Heck, Albert J.R.

AU - Schasfoort, Richardus B.M.

AU - Pruijn, Ger J.M.

N1 - Open access

PY - 2010

Y1 - 2010

N2 - Introduction Rheumatoid arthritis (RA) frequently involves the loss of tolerance to citrullinated antigens, which may play a role in pathogenicity. Citrullinated fibrinogen is commonly found in inflamed synovial tissue and is a frequent target of autoantibodies in RA patients. To obtain insight into the B-cell response to citrullinated fibrinogen in RA, its autoepitopes were systematically mapped using a new methodology. Methods Human fibrinogen was citrullinated in vitro by peptidylarginine deiminases (PAD), subjected to proteolysis and the resulting peptides were fractionated by ion exchange chromatography. The peptide composition of the citrullinated peptide-containing fractions was determined by high resolution tandem mass spectrometry. The recognition of these fractions by patient sera was subsequently analyzed by imaging surface plasmon resonance on microarrays. Results In total about two-thirds of the 81 arginines of human fibrinogen were found to be susceptible to citrullination by the human PAD2, the human PAD4 or the rabbit PAD2 enzymes. Citrullination sites were found in all three polypeptide chains of fibrinogen, although the α-chain appeared to contain most of them. The analysis of 98 anti-citrullinated protein antibody-positive RA sera using the new methodology allowed the identification of three major citrullinated epitope regions in human fibrinogen, two in the α- and one in the β-chain. Conclusions A comprehensive overview of citrullination sites in human fibrinogen was generated. The multiplex analysis of peptide fractions derived from a post-translationally modified protein, characterized by mass spectrometry, with patient sera provides a versatile system for mapping modified amino acid-containing epitopes. The citrullinated epitopes of human fibrinogen most efficiently recognized by RA autoantibodies are confined to three regions of its polypeptides.

AB - Introduction Rheumatoid arthritis (RA) frequently involves the loss of tolerance to citrullinated antigens, which may play a role in pathogenicity. Citrullinated fibrinogen is commonly found in inflamed synovial tissue and is a frequent target of autoantibodies in RA patients. To obtain insight into the B-cell response to citrullinated fibrinogen in RA, its autoepitopes were systematically mapped using a new methodology. Methods Human fibrinogen was citrullinated in vitro by peptidylarginine deiminases (PAD), subjected to proteolysis and the resulting peptides were fractionated by ion exchange chromatography. The peptide composition of the citrullinated peptide-containing fractions was determined by high resolution tandem mass spectrometry. The recognition of these fractions by patient sera was subsequently analyzed by imaging surface plasmon resonance on microarrays. Results In total about two-thirds of the 81 arginines of human fibrinogen were found to be susceptible to citrullination by the human PAD2, the human PAD4 or the rabbit PAD2 enzymes. Citrullination sites were found in all three polypeptide chains of fibrinogen, although the α-chain appeared to contain most of them. The analysis of 98 anti-citrullinated protein antibody-positive RA sera using the new methodology allowed the identification of three major citrullinated epitope regions in human fibrinogen, two in the α- and one in the β-chain. Conclusions A comprehensive overview of citrullination sites in human fibrinogen was generated. The multiplex analysis of peptide fractions derived from a post-translationally modified protein, characterized by mass spectrometry, with patient sera provides a versatile system for mapping modified amino acid-containing epitopes. The citrullinated epitopes of human fibrinogen most efficiently recognized by RA autoantibodies are confined to three regions of its polypeptides.

KW - METIS-282487

KW - IR-104395

U2 - 10.1186/ar3205

DO - 10.1186/ar3205

M3 - Article

VL - 12

SP - -

JO - Arthritis research & therapy

JF - Arthritis research & therapy

SN - 1478-6354

IS - 6

M1 - R219

ER -

van Beers JJBC, Raijmakers R, Alexander L-E, Stammen-Vogelzangs J, Lokate AMC, Heck AJR et al. Mapping of citrullinated fibrinogen B-cell epitopes in rheumatoid arthritis by imaging surface plasmon resonance. Arthritis research & therapy. 2010;12(6):-. R219. https://doi.org/10.1186/ar3205