Abstract
The impact of protein arginine methylation on the regulation of immune functions is virtually unknown. Here, we apply a novel method—isomethionine methyl-SILAC—coupled with antibody-mediated arginine-methylated peptide enrichment to identify methylated peptides in human T cells by mass spectrometry. This approach allowed the identification of 2,502 arginine methylation sites from 1,257 tissue-specific and housekeeping proteins. We find that components of T cell antigen receptor signal machinery and several key transcription factors that regulate T cell fate determination are methylated on arginine. Moreover, we demonstrate changes in arginine methylation stoichiometry during cellular stimulation in a subset of proteins critical to T cell differentiation. Our data suggest that protein arginine methyltransferases exert key regulatory roles in T cell activation and differentiation, opening a new field of investigation in T cell biology.
Highlights
The impact of protein arginine methylation on the regulation of immune functions is virtually unknown
We demonstrated that arginine methylation stoichiometry changes during cell differentiation and show this to occur in mRNA splicing factors critical in T cell differentiation
In heavy methyl-SILAC, cells are labelled with L-Methionine or LMethionine-13CD3
Summary
The impact of protein arginine methylation on the regulation of immune functions is virtually unknown. We apply a novel method—isomethionine methyl-SILAC—coupled with antibody-mediated arginine-methylated peptide enrichment to identify methylated peptides in human T cells by mass spectrometry. This approach allowed the identification of 2,502 arginine methylation sites from 1,257 tissue-specific and housekeeping proteins. We demonstrate changes in arginine methylation stoichiometry during cellular stimulation in a subset of proteins critical to T cell differentiation. When applied to Jurkat T cells and TCR/ CD28-stimulated primary T cells, this comprehensive approach allowed us to identify the largest number of arginine methylation sites and proteins known to date implicating PRMT action in most, if not all cell functions, including TCR-proximal signalling and cell fate programs. We demonstrated that arginine methylation stoichiometry changes during cell differentiation and show this to occur in mRNA splicing factors critical in T cell differentiation
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
