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Immunoaffinity Enrichment and Mass Spectrometry Analysis of Protein Methylation
Protein methylation is a common posttranslational modification that mostly occurs on arginine and lysine residues. Arginine methylation has been reported to regulate RNA processing, gene transcription, DNA damage repair, protein translocation, and signal transduction. Lysine methylation is best know...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Biochemistry and Molecular Biology
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3879628/ https://www.ncbi.nlm.nih.gov/pubmed/24129315 http://dx.doi.org/10.1074/mcp.O113.027870 |
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author | Guo, Ailan Gu, Hongbo Zhou, Jing Mulhern, Daniel Wang, Yi Lee, Kimberly A. Yang, Vicky Aguiar, Mike Kornhauser, Jon Jia, Xiaoying Ren, Jianmin Beausoleil, Sean A. Silva, Jeffrey C. Vemulapalli, Vidyasiri Bedford, Mark T. Comb, Michael J. |
author_facet | Guo, Ailan Gu, Hongbo Zhou, Jing Mulhern, Daniel Wang, Yi Lee, Kimberly A. Yang, Vicky Aguiar, Mike Kornhauser, Jon Jia, Xiaoying Ren, Jianmin Beausoleil, Sean A. Silva, Jeffrey C. Vemulapalli, Vidyasiri Bedford, Mark T. Comb, Michael J. |
author_sort | Guo, Ailan |
collection | PubMed |
description | Protein methylation is a common posttranslational modification that mostly occurs on arginine and lysine residues. Arginine methylation has been reported to regulate RNA processing, gene transcription, DNA damage repair, protein translocation, and signal transduction. Lysine methylation is best known to regulate histone function and is involved in epigenetic regulation of gene transcription. To better study protein methylation, we have developed highly specific antibodies against monomethyl arginine; asymmetric dimethyl arginine; and monomethyl, dimethyl, and trimethyl lysine motifs. These antibodies were used to perform immunoaffinity purification of methyl peptides followed by LC-MS/MS analysis to identify and quantify arginine and lysine methylation sites in several model studies. Overall, we identified over 1000 arginine methylation sites in human cell line and mouse tissues, and ∼160 lysine methylation sites in human cell line HCT116. The number of methylation sites identified in this study exceeds those found in the literature to date. Detailed analysis of arginine-methylated proteins observed in mouse brain compared with those found in mouse embryo shows a tissue-specific distribution of arginine methylation, and extends the types of proteins that are known to be arginine methylated to include many new protein types. Many arginine-methylated proteins that we identified from the brain, including receptors, ion channels, transporters, and vesicle proteins, are involved in synaptic transmission, whereas the most abundant methylated proteins identified from mouse embryo are transcriptional regulators and RNA processing proteins. |
format | Online Article Text |
id | pubmed-3879628 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | The American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-38796282014-01-13 Immunoaffinity Enrichment and Mass Spectrometry Analysis of Protein Methylation Guo, Ailan Gu, Hongbo Zhou, Jing Mulhern, Daniel Wang, Yi Lee, Kimberly A. Yang, Vicky Aguiar, Mike Kornhauser, Jon Jia, Xiaoying Ren, Jianmin Beausoleil, Sean A. Silva, Jeffrey C. Vemulapalli, Vidyasiri Bedford, Mark T. Comb, Michael J. Mol Cell Proteomics Technological Innovation and Resources Protein methylation is a common posttranslational modification that mostly occurs on arginine and lysine residues. Arginine methylation has been reported to regulate RNA processing, gene transcription, DNA damage repair, protein translocation, and signal transduction. Lysine methylation is best known to regulate histone function and is involved in epigenetic regulation of gene transcription. To better study protein methylation, we have developed highly specific antibodies against monomethyl arginine; asymmetric dimethyl arginine; and monomethyl, dimethyl, and trimethyl lysine motifs. These antibodies were used to perform immunoaffinity purification of methyl peptides followed by LC-MS/MS analysis to identify and quantify arginine and lysine methylation sites in several model studies. Overall, we identified over 1000 arginine methylation sites in human cell line and mouse tissues, and ∼160 lysine methylation sites in human cell line HCT116. The number of methylation sites identified in this study exceeds those found in the literature to date. Detailed analysis of arginine-methylated proteins observed in mouse brain compared with those found in mouse embryo shows a tissue-specific distribution of arginine methylation, and extends the types of proteins that are known to be arginine methylated to include many new protein types. Many arginine-methylated proteins that we identified from the brain, including receptors, ion channels, transporters, and vesicle proteins, are involved in synaptic transmission, whereas the most abundant methylated proteins identified from mouse embryo are transcriptional regulators and RNA processing proteins. The American Society for Biochemistry and Molecular Biology 2014-01 2013-10-15 /pmc/articles/PMC3879628/ /pubmed/24129315 http://dx.doi.org/10.1074/mcp.O113.027870 Text en © 2014 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. |
spellingShingle | Technological Innovation and Resources Guo, Ailan Gu, Hongbo Zhou, Jing Mulhern, Daniel Wang, Yi Lee, Kimberly A. Yang, Vicky Aguiar, Mike Kornhauser, Jon Jia, Xiaoying Ren, Jianmin Beausoleil, Sean A. Silva, Jeffrey C. Vemulapalli, Vidyasiri Bedford, Mark T. Comb, Michael J. Immunoaffinity Enrichment and Mass Spectrometry Analysis of Protein Methylation |
title | Immunoaffinity Enrichment and Mass Spectrometry Analysis of Protein Methylation |
title_full | Immunoaffinity Enrichment and Mass Spectrometry Analysis of Protein Methylation |
title_fullStr | Immunoaffinity Enrichment and Mass Spectrometry Analysis of Protein Methylation |
title_full_unstemmed | Immunoaffinity Enrichment and Mass Spectrometry Analysis of Protein Methylation |
title_short | Immunoaffinity Enrichment and Mass Spectrometry Analysis of Protein Methylation |
title_sort | immunoaffinity enrichment and mass spectrometry analysis of protein methylation |
topic | Technological Innovation and Resources |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3879628/ https://www.ncbi.nlm.nih.gov/pubmed/24129315 http://dx.doi.org/10.1074/mcp.O113.027870 |
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