Methylation

Introduction
Protein methylation is a post-translational modification process, in which proteins are modified
by an addition of methyl groups by S-adenosylmethionine dependent methyltransferases, with
S-adenosyl methionine (SAM) as the primary donor of methyl group. Methylation of proteins
can occur on multiple amino acids on proteins, including arginine, lysine, histidine, etc.
Methylation is mediated by methyltransferases, Methylation of lysine involves the addition of
one to three methyl groups on the amino acid's ε-amine group, to form mono-, di- or tri-
methyllysine. This modification often occurs in a conserved sequence motif on proteins. For
example, two typical motifs are RGG box and RXR sequence. Depending on the site of
residual methyl group attached to, there are two types of methylation: N-methylation and O-
methylation. N-methylation is irreversible, whereas, O-methylation is potentially reversible.

The transfer of one-carbon methyl groups to nitrogen or oxygen (N- and O-methylation,
respectively) to amino acid side chains increases the hydrophobicity of the protein and can
neutralize a negative amino acid charge when bound to carboxylic acids. Methylation on the
tails of histone proteins, in conjunction with acetylation and phosphorylation, controls their
interaction with other proteins, affects chromatin compaction and the up- or down-regulation
of gene expression.

Creative Proteomics has established a highly sensitive HPLC-MS/MS pipeline that can
analyze N- and O-methylation in both eukaryotic and prokaryotic organisms. In addition, we
have optimized our protocol, to enable more fast and sensitive site mapping service for
methylation analysis.
 Workflow of our Methylation analysis service:
 In gel or in solution digestion of proteins
 Enrichment of methylated proteins by antibodies targeting specific methylation motifs
(optional step)
 HPLC separation, followed by ESI-TOF MS/MS analysis
 Mass spectrometry data interpretation
Technology platform:
 Ion Chromatography
 High Performance Liquid Chromatography (HPLC)
 Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI-MS)
Creative Proteomics also provide the following bioinformatics services in Protein Post-
translational Modification Analysis:
Functional annotation and enrichment analysis
Clustering analysis
Network analysis
Statistical analysis
Proteomic analysis of post-translational modifications
Please feel free to Contact Us to discuss your projects. We hope you will find that we can
meet your research needs.