Using proximity ligation assay to detect protein arginine methylation

Methods. 2020 Mar 15;175:66-71. doi: 10.1016/j.ymeth.2019.09.007. Epub 2019 Sep 6.


Arginine methylation is now recognized as a major contributor to proteome diversity and is, as such, involved in a large range of cellular processes. There is a growing need for assessing endogenous protein arginine methylation in cells. Besides the classical immunoprecipitation, in situ proximity ligation assay (PLA) is a useful technique allowing at the same time the detection, localization and quantification of arginine methylation of a given protein within a cellular context. Here, we described in depth a standard PLA protocol applied to the detection of arginine methylation in combination with RNA interference and specific methyltransferase inhibitors. We demonstrated that the glucocorticoid receptor is methylated by the arginine methyltransferase PRMT5 inside the nucleus of MCF-7 cells. In addition, the automated quantification of protein arginine methylation performed using Image J is reported. Hence, we demonstrated that PLA offers a novel approach to study protein arginine methylation and could be extended to other post-translational modifications when specific antibodies are available.

Keywords: Arginine methylation; Glucocorticoid receptor; PRMT5; Proximity ligation assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arginine / metabolism*
  • Cell Nucleus / metabolism
  • DNA Ligases / chemistry
  • Enzyme Assays / methods*
  • Enzyme Inhibitors / chemistry
  • Epigenomics / methods*
  • Humans
  • Immunoprecipitation
  • MCF-7 Cells
  • Methylation
  • Protein Processing, Post-Translational*
  • Protein-Arginine N-Methyltransferases / antagonists & inhibitors
  • Protein-Arginine N-Methyltransferases / metabolism*
  • RNA Interference
  • Receptors, Glucocorticoid / metabolism*
  • Sensitivity and Specificity
  • Software


  • Enzyme Inhibitors
  • Receptors, Glucocorticoid
  • Arginine
  • PRMT5 protein, human
  • Protein-Arginine N-Methyltransferases
  • DNA Ligases