Citrullination of collagen II affects integrin-mediated cell adhesion in a receptor-specific manner.

Abstract

Citrullinated collagen II (CII) is a well-known autoantigen in rheumatoid arthritis (RA). However, the direct effects of CII citrullination on cell behavior have not been described. To study whether citrullination of CII could affect cellular functions, we measured the adhesion of 3 different cell types (human Saos2 osteosarcoma cells, human synovial fibroblasts, and rat mesenchymal stem cells) with impedance-based technology. The binding of different collagen receptor integrins to citrullinated collagen was studied by CHO cell lines, each overexpressing 1 of the 4 human collagen receptors on the cell surface, and with solid-phase binding assays, using the recombinant human integrin α1I, α2I, α10I, and α11I domains. Collagen citrullination decreased the adhesion of synovial fibroblasts ∼50% (P<0.05) and mesenchymal stem cells ∼40% (P<0.05) by specifically decreasing the binding of integrins α10β1 and α11β1 to arginine-containing motifs, such as GFOGER. In contrast, citrullination had only a minor effect on the function of α1β1 and α2β1 integrins, which have been reported to play a critical role in regulating leukocyte function. Molecular modeling was used to explain the detected functional differences at the structural level. Given that the integrins regulate cell metabolism, proliferation, and migration, we suggest that collagen citrullination modifies the pathogenesis of RA. Here, CII citrullination was shown to decrease the survival of mesenchymal stem cells.

• Authors: Kalle Sipilä, Sabrina Haag, Konstantin Denessiouk, Jarmo Käpylä, Eric C Peters, Alexander Denesyuk, Uwe Hansen, Yrjö Konttinen, Mark S Johnson, Rikard Holmdahl, Jyrki Heino
• Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology (FASEB J) Vol. 28 Issue 8 Pg. 3758-68 (Aug 2014) ISSN: 1530-6860 [Electronic] United States
• PMID: 24823363 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © FASEB.

Chemical References

• Collagen Type II
• Integrins
• Peptide Fragments
• Recombinant Fusion Proteins
• Citrulline
• Arginine

Topics

• Amino Acid Motifs
• Aminoacylation
• Animals
• Arginine (chemistry)
• Bone Neoplasms (pathology)
• CHO Cells
• Cell Adhesion (physiology)
• Cell Line, Tumor
• Cells, Cultured
• Chickens
• Citrulline (chemistry)
• Collagen Type II (chemistry, pharmacology)
• Cricetinae
• Cricetulus
• Fibroblasts (pathology)
• Humans
• Integrins (genetics, physiology)
• Mesenchymal Stem Cells (cytology, drug effects)
• Osteoarthritis (pathology)
• Osteosarcoma (pathology)
• Peptide Fragments (chemical synthesis, chemistry, metabolism)
• Protein Binding
• Protein Processing, Post-Translational
• Rats
• Recombinant Fusion Proteins (metabolism)
• Synovial Membrane (pathology)
• Transfection