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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...

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Autores principales: Uhlmann, Thomas, Geoghegan, Vincent L., Thomas, Benjamin, Ridlova, Gabriela, Trudgian, David C., Acuto, Oreste
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Biochemistry and Molecular Biology 2012
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.
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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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