Immunoaffinity enrichment and mass spectrometry analysis of protein methylation.

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Guo A, Gu H, Zhou J, Mulhern D, Wang Y, Lee KA, Yang V, Aguiar M, Kornhauser J, Jia X, Ren J, Beausoleil SA, Silva JC, Vemulapalli V, Bedford MT, Comb MJ

Immunoaffinity enrichment and mass spectrometry analysis of protein methylation.

Mol Cell Proteomics. 2014 Jan;13(1):372-87. doi: 10.1074/mcp.O113.027870. Epub 2013 Oct 15.

PubMed ID
24129315 [ View in PubMed
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Abstract

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 approximately 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.

DrugBank Data that Cites this Article

Polypeptides
NameUniProt ID
Calmodulin-2P0DP24Details
Calmodulin-3P0DP25Details
Heterogeneous nuclear ribonucleoproteins A2/B1P22626Details
Heterogeneous nuclear ribonucleoprotein HP31943Details
Heterogeneous nuclear ribonucleoprotein H3P31942Details
Splicing factor, proline- and glutamine-richP23246Details
Protein disulfide-isomerase A3P30101Details
Heterogeneous nuclear ribonucleoprotein KP61978Details
Heat shock 70 kDa protein 4P34932Details
SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily E member 1Q969G3Details
Far upstream element-binding protein 2Q92945Details
Heterogeneous nuclear ribonucleoprotein D0Q14103Details
Small ribosomal subunit protein eS19P39019Details
Splicing factor 1Q15637Details
Transgelin-2P37802Details
ZyxinQ15942Details
Serine/arginine-rich splicing factor 1Q07955Details
TransgelinQ01995Details
Mitogen-activated protein kinase 15Q8TD08Details
Misshapen-like kinase 1Q8N4C8Details
Serine/threonine-protein kinase Nek4P51957Details
Serine/threonine-protein kinase PAK 4O96013Details
Voltage-gated potassium channel subunit beta-2Q13303Details
ELAV-like protein 1Q15717Details
Dual specificity mitogen-activated protein kinase kinase 4P45985Details
Thioredoxin reductase 3Q99MD6Details