Protein methylation is a post-translational modification (PTM) process, in which highly specific enzymes called methyltransferases are responsible for the addition of methyl groups to a targeted molecule and S-adenosyl methionine (SAM) as the primary donor of methyl group. Protein methylation commonly occurs on arginine, lysine, histidine, proline, and carboxyl groups. Protein methylation plays an important role in modulating cellular and biological processes, including transcriptional regulation, RNA processing, metabolism and signal transduction.
Figure 1. Types of methylation on arginine residues (Yang Y and Bedford M T, 2013).
Overview of Protein Methylation Service
Creative Proteomics has already developed a highly sensitive HPLC-MS/MS pipeline that can analyze N- and O-methylation. With powerful and sensitive tools, we can identify, quantify and characterize protein methylation. Metabolic labeling strategies can be coupled with MS to measure dynamic and differential in vivo protein methylation rates. In addition, we also provide bioinformatics services in Protein Post-translational Modification Analysis. We have optimized our protocol to enable more fast and sensitive services for methylation analysis. Shown as Figure 2, our protein methylation service contains digestion, enrichment, LC-MS/MS analysis, and data analysis.
Figure 2. The workflow of protein methylation analysis.
Sample Requirements
- Plant roots, xylem, phloem, etc.: 5g or more
- Animal tissues: wet weight >200 mg
- Microorganisms: wet weight > 2 g
- Body fluids (saliva, amniotic fluid, cerebrospinal fluid, etc.): 10 mL or more
- Serum: 500 μL or more
- Urine: 50 mL or more
- Protein extract: concentration > 2 mg/mL, total not less than 1 mg. In order to ensure the test results, please inform the buffer components, whether it contains thiourea, SDS, or strong ion salts. In addition, the sample should not contain components such as nucleic acids, lipids, and polysaccharides, which will affect the separation effect.
If you want to know specific samples requirements, please feel free to contact us.
Delivery
- Detailed report, including experimental materials, methods, procedures, and results
- Raw data and data analysis result
Our Advantages
- High-throughput: Identify and quantify up to thousands of proteins at once
- 100% coverage: Use 2-3 enzymes for protein digestion to ensure full-coverage
- Ability to analyze low abundance ubiquitin-proteins
- State-of-the-art facilities: Triple TOF 5600, Q-Exactive, Orbitrap Fusion Tribrid
- Constantly optimized and validated protocol
Technology platforms
- Ion Chromatography
- High Performance Liquid Chromatography (HPLC)
- Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI-MS)
Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI-MS)
At Creative Proteomics, many excellent and experienced experts will optimize the experimental protocol according to your requirement and guarantee the high-quality results for protein methylation analysis. As every project has different requirements, please contact our specialists to discuss your specific needs.
Reference
1. Yang Y, Bedford M T. Protein arginine methyltransferases and cancer. Nature Reviews Cancer, 2013, 13(1): 37.