Methylproteomics Analysis Service
- Sample Compatibility: Suitable for fresh/frozen tissues, cell lysates, serum/plasma, and other biofluids.
- Methylation Enrichment: Selection of antibody-based or chemical enrichment strategies (e.g., strong cation exchange chromatography) for arginine- and lysine-specific modifications, enhancing target peptide recovery.
- LC-MS/MS Platform: High-resolution Orbitrap-based mass spectrometry supporting both untargeted (DDA) and targeted (DIA) analysis.
- Database Matching: Reliable identification using UniProt and other curated databases, with manual validation of methylation sites.
- Bioinformatics Analysis: Differential methylation profiling; Pathway enrichment analysis; Interaction network construction; Functional annotation
- Comprehensive Methylation Site List-Including modification types, localization probability, and abundance information.
- Functional Annotation and Mechanistic Insights-Detailed analysis of methylation-related biological processes.
- Raw Data and Editable Graphical Outputs-Allowing further customized analysis.
- Optional Integrative Analysis-Multi-omics integration (e.g., methylation-phosphorylation or methylation-acetylation cross-talk analysis).
- Optimized antibody enrichment workflows and low-input detection capabilities for reliable profiling of trace-level methylation.
- Stringent quality control ensuring >95% site identification accuracy.
- Custom enrichment and detection strategies based on specific methylation types (e.g., asymmetric dimethyl-arginine).
- Our platform is compatible with human, mouse, rat, plant, and other model organisms.
- Standard projects completed within 3-6 weeks.
- We support pilot studies for small-scale experiments and large-cohort studies for clinical research.
- Biomarker Discovery: Identifying methylation signatures from biofluids or tissues linked to disease staging and prognosis, supporting non-invasive diagnostic tool development.
- Therapeutic Target Identification: Discovering methylation-modified functional proteins and regulatory enzymes, providing a basis for targeted drug development.
Protein methylation is a key post-translational modification (PTM) involving the covalent addition of methyl groups to specific residues, predominantly arginine (Arg) and lysine (Lys). This dynamic modification plays a crucial role in gene expression regulation, signal transduction, RNA metabolism, and disease pathogenesis. Methylproteomics aims to comprehensively characterize methylation sites, modification abundance, and biological significance, providing essential insights for disease mechanism research, biomarker discovery, and drug target identification. However, due to the low abundance, wide dynamic range, and lack of specific antibodies for methylation, achieving high-sensitivity detection requires advanced analytical platforms.
Zhong, Q. et al. MedComm (2020). 2023.
Figure 1. Functions of Protein Methylation
At MtoZ Biolabs, we leverage cutting-edge LC-MS/MS platforms combined with targeted enrichment strategies (including high-specificity antibodies and chemical capture techniques) to offer one-stop methylproteomics analysis service. Our rigorous experimental design and specialized analytical workflows enable researchers to overcome technical challenges and achieve precise characterization of methylation dynamics, facilitating scientific discovery and translational applications.
Our service provides comprehensive identification and quantification of protein methylation modifications across various biological samples (tissues, cells, biofluids). We cover multiple arginine methylation types (monomethylation, asymmetric/symmetric dimethylation) and lysine methylation states (mono-, di-, and trimethylation).
By integrating multi-dimensional enrichment strategies with high-resolution mass spectrometry, we enable:
1. High-Sensitivity Detection-Efficient capture and identification of low-abundance methylated peptides.
2. Accurate Site Localization-Verification via spectral analysis and sequence alignment for precise methylation site identification.
3. Targeted Quantification-Comparative methylation level analysis across different sample conditions.
4. Functional Correlation Analysis-Integration of bioinformatics tools to uncover associations between methylation and disease pathways.
Analysis Workflow
1. Sample Preparation and Methylation Peptide Enrichment
2. High-Precision Mass Spectrometry and Data Analysis
3. Professional Data Reporting
Pascovici, D. et al. Int J Mol Sci. 2018.
Figure 2. The Workflow of Protein PTM Analysis
Why Choose MtoZ Biolabs?
1. Technological Expertise
2. Specialized Analytical Support
3. Efficient Turnaround Time
4. Expertise in Complex Sample Handling
Extensive experience with challenging sample types (e.g., brain tissue, exosomes, and clinical plasma).
Applications
1. Disease Mechanism Exploration
Protein methylation plays a pivotal role in cell signaling, metabolic reprogramming, and pathological microenvironment remodeling. Global methylproteomics analysis enables researchers to uncover disease-associated methylation networks, identifying key regulatory nodes that drive disease progression, such as dysregulated methyltransferase activity or aberrant signaling pathways.
2. Translational Medicine Research
3. Precision Medicine Applications
Methylation heterogeneity correlates with individual phenotypic variations. Large-scale methylation profiling enables patient stratification, guiding personalized treatment strategies for improved therapeutic outcomes.
Case Study
1. Antibody-Free Approach for the Global Analysis of Protein Methylation
Protein methylation plays an important role in various biological processes, including gene transcription regulation, RNA processing, DNA damage repair, and signal transduction. However, due to the lack of broad-specificity antibodies, immunoaffinity enrichment methods face difficulties in achieving a comprehensive proteomic analysis of protein methylation. This study developed an antibody-free enrichment method based on the principle that lysine (K) and arginine (R) methylation prevent protease cleavage. Using this method, 887 methylation forms covering 768 sites were identified from HepG2 cells, significantly enhancing the detection of arginine methylation. This method enables the simultaneous analysis of both lysine and arginine methylation, providing a powerful tool for studying the biological processes regulated by methylation. Methylproteomics Analysis Service utilizes high-resolution mass spectrometry technology combined with optimized enrichment strategies to provide comprehensive analysis of protein methylation. It can precisely identify lysine and arginine methylation sites, analyze their dynamic changes and biological regulatory roles, and help explore the role of methylation in cellular functions and signaling pathways. The service is suitable for studying the biological processes driven by methylation modifications and their regulatory mechanisms.
Wang, K. et al. Anal Chem. 2016.
Figure 3. The Global View of Methylproteome of HepG2 Cells
2. Quantitative Analysis of the Protein Methylome Reveals PARP1 Methylation is involved in DNA Damage Response
This study conducted quantitative analysis of protein methylation in HEK293T cells under ionizing radiation (IR) treatment using HILIC affinity enrichment combined with mass spectrometry. A total of 235 methylation sites were identified, 38% of which were newly discovered. Multiple RNA-binding proteins exhibited differential methylation upon DNA damage stress, and 14 novel methylation sites were found in DNA damage response-related proteins. Furthermore, the study validated the function of PARP1 K23 methylation in repairing IR-induced DNA lesions, where the loss of this modification sensitized cancer cells to radiation and replication stress, while also affecting PARP1 binding to stalled replication forks. Methylproteomics Analysis Service utilizes high-resolution mass spectrometry combined with optimized enrichment strategies to enable comprehensive quantitative analysis of protein methylation. It provides insights into the dynamic changes of lysine and arginine methylation in DNA damage response, signaling pathways, and cellular regulation, facilitating the exploration of key methylation modifications in biological processes.
Wang, X. et al. Front Mol Biosci. 2022.
Figure 4. Subcellular localization and Functional Annotation of Identified Methylated Proteins upon IR Treatment
Methylproteomics is at the forefront of disease mechanism elucidation and precision medicine. However, its complexity requires advanced analytical platforms and expert data interpretation. At MtoZ Biolabs, we offer state-of-the-art technology, customizable solutions, and high-efficiency workflows to support your methylation research. Whether your focus is on deciphering fundamental methylation regulatory networks or developing clinical biomarkers and translational applications, we provide end-to-end solutions to help you decode the biological significance of methylation dynamics. Contact the MtoZ Biolabs team today to initiate your methylproteomics research journey!
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