Applications and Advantages of Mass Spectrometry in Post-Translational Modifications Analysis
-
Modification sites are typically low in abundance and susceptible to degradation;
-
Multiple types of modifications often coexist, with diverse structural features;
-
Precise identification of both the site and type of modification is required.
-
A workflow library for enriching diverse types of modifications, including phosphorylation, acetylation, ubiquitination, methylation, glycosylation, and SUMOylation;
-
A suite of high-resolution mass spectrometry systems supporting various detection modes, such as DDA, DIA, PRM, and MS³;
-
Capacity for large-scale project support, including clinical cohorts, integrative multi-omics analyses, and big data-driven investigations;
-
Comprehensive, one-stop deliverables including site-specific modification data, quantitative protein profiles, pathway enrichment analyses, and visualized charts;
-
Tailored scientific support, offering publication-ready data, project design consultation, and collaborative technology development.
Post-translational modifications (PTMs) are covalent enzymatic alterations that occur after protein translation, including phosphorylation, acetylation, ubiquitination, and methylation. These modifications regulate protein activity, subcellular localization, molecular interactions, and degradation. They represent essential molecular mechanisms for maintaining cellular homeostasis and for responding to environmental stimuli. However, the inherent complexity and dynamic nature of PTMs present significant challenges for analytical detection:
Among various analytical techniques, mass spectrometry (MS) has emerged as a pivotal tool for PTM analysis, owing to its high sensitivity, throughput, and resolution. In particular, when coupled with efficient enrichment strategies for modified peptides, MS can comprehensively address the analytical demands associated with most major types of PTMs.
Advantages of Using Mass Spectrometry for PTM Analysis
1. Ionization and Fragmentation: Identification of Modifications at the Peptide Level
Mass spectrometry enables the identification of peptide modifications through ionization, mass analysis, and tandem mass spectrometry (MS/MS). This approach allows for accurate determination of peptide mass changes and precise localization of modification sites.
(1) Precursor ion (MS1): reveals whether a peptide is modified based on observed mass shifts.
(2) Fragment ions (MS2): enable pinpoint localization of the modified amino acid residue.
2. Advantages of High-Resolution Mass Spectrometry Platforms
Leading high-resolution MS platforms such as Orbitrap, timsTOF, and Q-TOF systems offer the following advantages:
MtoZ Biolabs is equipped with Orbitrap Exploris 480 and Bruker timsTOF Pro systems, which support comprehensive detection of post-translational modifications, ranging from common types to specific low-abundance events such as SUMOylation and hydroxylation.
Application Practices of Mass Spectrometry in Mainstream PTM Types
1. Phosphorylation
(1) Enrichment strategies: TiO₂, IMAC, and Fe³⁺ magnetic beads;
(2) Challenges in MS detection: phosphorylation is labile and prone to neutral loss during ionization;
(3) Solution: application of HCD or ETD fragmentation techniques to preserve phosphate groups;
By optimizing a TiO₂-based enrichment combined with DDA/DIA acquisition platforms, MtoZ Biolabs consistently quantifies over 10,000 phosphorylation sites in representative tumor cell samples.
2. Acetylation & Methylation
(1) Enrichment strategies: enrichment using modification-specific antibodies (anti-AcK, anti-MeR);
(2) Quantification approaches: TMT labeling or label-free quantification;
(3) Biological relevance: commonly employed in the study of chromatin architecture, transcriptional regulation, and histone modifications;
3. Ubiquitination
(1) Signature motif: K-ε-GG residues specifically enriched;
(2) MS spectral features: diagnostic ions in MS2 spectra allow for determination of ubiquitin linkage types (e.g., K48, K63);
(3) Research applications: identification of degradation signals and functional characterization of E3 ubiquitin ligases.
Summary of the Unique Advantages of Mass Spectrometry in PTM Research
Service Capabilities of MtoZ Biolabs in PTM Mass Spectrometry Research
MtoZ Biolabs has cultivated deep expertise in mass spectrometry and proteomics, establishing a comprehensive platform for PTM analysis with the following capabilities:
As a central mechanism regulating protein function, post-translational modifications (PTMs) offer essential insights into complex biological systems. The application of mass spectrometry has introduced unprecedented resolution and systemic analysis capabilities in this field. It not only enables the detection of trace-level modifications but also reveals their spatial and temporal dynamics. This has transformed PTM research from isolated identification to system-wide quantification. MtoZ Biolabs continues to enhance its mass spectrometry platforms, optimize modification enrichment strategies, and standardize data analysis workflows to support researchers in elucidating PTM networks and advancing precision medicine and translational research.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
Related Services
How to order?
