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1

Department of Biomolecular Chemistry, Institute for Molecules and Materials, Radboud Institute for Molecular Life Sciences and Netherlands Proteomics Centre, Radboud University Nijmegen, P.O. Box 9101, NL-6500 HB Nijmegen, The Netherlands

2

Department of Synthetic Organic Chemistry, Institute for Molecules and Materials, Radboud University Nijmegen, P.O. Box 9101, NL-6500 HB Nijmegen, The Netherlands





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Academic Editor: Arnaud Gautier

Abstract Citrullination is the conversion of peptidylarginine to peptidylcitrulline, which is catalyzed by peptidylarginine deiminases. This conversion is involved in different physiological processes and is associated with several diseases, including cancer and rheumatoid arthritis. A common method to detect citrullinated proteins relies on anti-modified citrulline antibodies directed to a specific chemical modification of the citrulline side chain. Here, we describe a versatile, antibody-independent method for the detection of citrullinated proteins on a membrane, based on the selective reaction of phenylglyoxal with the ureido group of citrulline under highly acidic conditions. The method makes use of 4-azidophenylglyoxal, which, after reaction with citrullinated proteins, can be visualized with alkyne-conjugated probes. The sensitivity of this procedure, using an alkyne-biotin probe, appeared to be comparable to the antibody-based detection method and independent of the sequence surrounding the citrulline. View Full-Text

Keywords: citrulline; peptidylarginine deiminases; anti-modified citrulline antibodies; phenylglyoxal; azide-alkyne cycloaddition; rheumatoid arthritis citrulline; peptidylarginine deiminases; anti-modified citrulline antibodies; phenylglyoxal; azide-alkyne cycloaddition; rheumatoid arthritis





Author: Sanne M. M. Hensen 1, Wilbert C. Boelens 1, Kimberly M. Bonger 1, Remco T. P. van Cruchten 1, Floris L. van Delft 2 and Ger J. M. Pruijn 1,*

Source: http://mdpi.com/



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