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Identification of novel citrullinated proteins in Rheumatoid Arthritis patients

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Identification of novel citrullinated proteins in Rheumatoid Arthritis patients

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Samenvatting

Rheumatoid Arthritis (RA) is a common autoimmune disease. The disease is characterized by inflammation and swelling of the synovial joints. Specific for RA is the presence of autoantibodies against citrullinated proteins. The role of these citrullinated proteins in RA pathogenesis still remains unclear. Research on citrullinated proteins and their antigenic properties may give more insight in the molecular mechanism behind RA and may contribute to a better understanding of the cause and course of the disease. Furthermore, citrullinated proteins have proven to be useful in the detection of their autoantibodies in patients. Citrullinated peptides are used in the clinic nowadays for diagnosing RA. Newly identified citrullinated proteins can contribute to a better understanding of the disease and to diagnostic improvements. To identify new citrullinated proteins in RA patients, synovial fluids were analyzed by TANDEM mass spectrometry (MS). Synovial fluids of 33 RA patients were screened for the presence of citrullinated proteins by separation on a SDS-PAGE gel and detection on western blot with an anti-modified citrulline antibody. Thirteen synovial fluids containing citrullinated proteins were depleted for the abundant protein albumin. Albumin depleted samples were separated by size on a SDS-PAGE gel. The gel was stained with colloidal coomassie (CBB) staining and each sample lane was cut into bands. A matching western blot was made with anti-modified citrulline antibody to compare and determine which cut out bands from the CBB stained gel contained citrullinated proteins. From each citrullinated protein containing gel band a protein fragment mix was obtained by in-gel digestion with trypsin. The protein fragment mixes were analyzed by TANDEM MS. As a result 57 citrullinated proteins were identified. Five of the identified proteins are already known citrullinated proteins in RA patients. Eight citrullinated proteins were identified that contained multiple citrulline residues. Literature pointed out that these proteins may have functions involved in RA/ inflammatory related processes. Therefore these citrullinated proteins were marked as possible candidate auto-antigens.

Toon meer
OrganisatieHogeschool Utrecht
OpleidingBiologie en Medisch Laboratorium Onderzoek
AfdelingLife Sciences en Chemistry
PartnersNijmegen Centre for Molecular Life Sciences Department of Biomolecular Chemistry
Jaar2007
TypeBachelor
TaalEngels

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