Phenylglyoxal-Based Visualization of Citrullinated Proteins on Western Blots
AbstractCitrullination 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
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Hensen, S.M.M.; Boelens, W.C.; Bonger, K.M.; van Cruchten, R.T.P.; van Delft, F.L.; Pruijn, G.J.M. Phenylglyoxal-Based Visualization of Citrullinated Proteins on Western Blots. Molecules 2015, 20, 6592-6600.
Hensen SMM, Boelens WC, Bonger KM, van Cruchten RTP, van Delft FL, Pruijn GJM. Phenylglyoxal-Based Visualization of Citrullinated Proteins on Western Blots. Molecules. 2015; 20(4):6592-6600.Chicago/Turabian Style
Hensen, Sanne M.M.; Boelens, Wilbert C.; Bonger, Kimberly M.; van Cruchten, Remco T.P.; van Delft, Floris L.; Pruijn, Ger J.M. 2015. "Phenylglyoxal-Based Visualization of Citrullinated Proteins on Western Blots." Molecules 20, no. 4: 6592-6600.