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A Method for Large-scale Identification of Protein Arginine Methylation
The lack of methods for proteome-scale detection of arginine methylation restricts our knowledge of its relevance in physiological and pathological processes. Here we show that most tryptic peptides containing methylated arginine(s) are highly basic and hydrophilic. Consequently, they could be consi...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Biochemistry and Molecular Biology
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3494207/ https://www.ncbi.nlm.nih.gov/pubmed/22865923 http://dx.doi.org/10.1074/mcp.M112.020743 |
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author | Uhlmann, Thomas Geoghegan, Vincent L. Thomas, Benjamin Ridlova, Gabriela Trudgian, David C. Acuto, Oreste |
author_facet | Uhlmann, Thomas Geoghegan, Vincent L. Thomas, Benjamin Ridlova, Gabriela Trudgian, David C. Acuto, Oreste |
author_sort | Uhlmann, Thomas |
collection | PubMed |
description | The lack of methods for proteome-scale detection of arginine methylation restricts our knowledge of its relevance in physiological and pathological processes. Here we show that most tryptic peptides containing methylated arginine(s) are highly basic and hydrophilic. Consequently, they could be considerably enriched from total cell extracts by simple protocols using either one of strong cation exchange chromatography, isoelectric focusing, or hydrophilic interaction liquid chromatography, the latter being by far the most effective of all. These methods, coupled with heavy methyl-stable isotope labeling by amino acids in cell culture and mass spectrometry, enabled in T cells the identification of 249 arginine methylation sites in 131 proteins, including 190 new sites and 93 proteins not previously known to be arginine methylated. By extending considerably the number of known arginine methylation sites, our data reveal a novel proline-rich consensus motif and identify for the first time arginine methylation in proteins involved in cytoskeleton rearrangement at the immunological synapse and in endosomal trafficking. |
format | Online Article Text |
id | pubmed-3494207 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | The American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-34942072012-11-09 A Method for Large-scale Identification of Protein Arginine Methylation Uhlmann, Thomas Geoghegan, Vincent L. Thomas, Benjamin Ridlova, Gabriela Trudgian, David C. Acuto, Oreste Mol Cell Proteomics Technological Innovation and Resources The lack of methods for proteome-scale detection of arginine methylation restricts our knowledge of its relevance in physiological and pathological processes. Here we show that most tryptic peptides containing methylated arginine(s) are highly basic and hydrophilic. Consequently, they could be considerably enriched from total cell extracts by simple protocols using either one of strong cation exchange chromatography, isoelectric focusing, or hydrophilic interaction liquid chromatography, the latter being by far the most effective of all. These methods, coupled with heavy methyl-stable isotope labeling by amino acids in cell culture and mass spectrometry, enabled in T cells the identification of 249 arginine methylation sites in 131 proteins, including 190 new sites and 93 proteins not previously known to be arginine methylated. By extending considerably the number of known arginine methylation sites, our data reveal a novel proline-rich consensus motif and identify for the first time arginine methylation in proteins involved in cytoskeleton rearrangement at the immunological synapse and in endosomal trafficking. The American Society for Biochemistry and Molecular Biology 2012-11 2012-08-03 /pmc/articles/PMC3494207/ /pubmed/22865923 http://dx.doi.org/10.1074/mcp.M112.020743 Text en © 2012 by The American Society for Biochemistry and Molecular Biology, Inc. Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) applies to Author Choice Articles |
spellingShingle | Technological Innovation and Resources Uhlmann, Thomas Geoghegan, Vincent L. Thomas, Benjamin Ridlova, Gabriela Trudgian, David C. Acuto, Oreste A Method for Large-scale Identification of Protein Arginine Methylation |
title | A Method for Large-scale Identification of Protein Arginine Methylation |
title_full | A Method for Large-scale Identification of Protein Arginine Methylation |
title_fullStr | A Method for Large-scale Identification of Protein Arginine Methylation |
title_full_unstemmed | A Method for Large-scale Identification of Protein Arginine Methylation |
title_short | A Method for Large-scale Identification of Protein Arginine Methylation |
title_sort | method for large-scale identification of protein arginine methylation |
topic | Technological Innovation and Resources |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3494207/ https://www.ncbi.nlm.nih.gov/pubmed/22865923 http://dx.doi.org/10.1074/mcp.M112.020743 |
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