Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Biochemistry and Molecular Biology 2014
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
_version_ 1782298010424705024
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
work_keys_str_mv AT guoailan immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT guhongbo immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT zhoujing immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT mulherndaniel immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT wangyi immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT leekimberlya immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT yangvicky immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT aguiarmike immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT kornhauserjon immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT jiaxiaoying immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT renjianmin immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT beausoleilseana immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT silvajeffreyc immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT vemulapallividyasiri immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT bedfordmarkt immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation
AT combmichaelj immunoaffinityenrichmentandmassspectrometryanalysisofproteinmethylation