A highly selective electrochemical assay based on the Sakaguchi reaction for the detection of protein arginine methylation state

Abstract

Protein arginine methylation is a common form of post-translational modification that plays an important role in many bioprocesses. However, research advances in this field have been severely hampered by the lack of a quick and sensitive method for detecting the arginine methylation state of a protein. In this work we propose a direct and sensitive electrochemical method for identifying the arginine methylation state. This novel assay combines an electrochemical technique with the Sakaguchi reaction, which is highly selective towards the arginine methylation state. We show that the presence of a methyl group on the arginine residue of a protein prevents the Sakaguchi reaction, while the unmethylated arginine residue selectively reacts with 8-hydroxyquinoline; the electrical signal of the reaction product is used for electrochemical detection. From this, a highly selective and simple electrochemical sensor has been developed based on (1) the high selectivity of the Sakaguchi reaction towards the arginine methylation state, and (2) the sensitive electrochemical signal generated by the linked 8-hydroxyquinoline. The assay described in this work thus provides a convenient tool for detection of protein arginine methylation, which may facilitate studies of the biological functions of protein arginine methylases and demethylases.

Keywords

• Electrochemical assay
• Protein arginine methylation
• Sakaguchi reaction
• Biosensor