Crotonylation Proteomics Analysis Service
- Epigenetics: Investigating the role of crotonylation in histone modification and gene expression regulation.
- Cancer Biology: Profiling crotonylation patterns in tumor-related proteins to identify potential biomarkers and therapeutic targets.
- Metabolic Regulation: Exploring the involvement of crotonylation in glucose and lipid metabolism.
- Neuroscience: Examining crotonylation in neuronal proteins to understand its impact on synaptic plasticity and neurodegenerative diseases.
- Immunology: Analyzing how crotonylation influences immune response pathways and immune cell function.
Crotonylation proteomics analysis is a mass spectrometry-based approach dedicated to identifying and quantifying lysine crotonylation modifications in proteins. First discovered in 2011, lysine crotonylation is a key post-translational modification (PTM) widely present in eukaryotic cells, particularly in histones, where it plays a critical role in gene expression regulation, chromatin remodeling, and cellular metabolism.
Advancements in epigenetics and PTM research have revealed that lysine crotonylation extends beyond nuclear proteins, being prevalent in cytoplasmic, mitochondrial, and signaling pathway proteins. It is implicated in various biological processes, including energy metabolism, cell proliferation, tumorigenesis, and immune regulation. A systematic analysis of crotonylated proteomes provides insights into its functional mechanisms across different physiological and pathological conditions.
Figure 1. Lysine Crotonylation
MtoZ Biolabs offers high-sensitivity, high-resolution Crotonylation Proteomics Analysis Service that integrates antibody enrichment techniques (Antibody Enrichment) with liquid chromatography-tandem mass spectrometry (LC-MS/MS). This service enables comprehensive analysis of the distribution, abundance variations, and potential biological functions of crotonylation modification.
Technical Principles
Crotonylation is a dynamic and reversible post-translational modification that increases the molecular weight of lysine residues by 68.0262 Da, producing a distinct mass shift signature in mass spectrometry detection. MtoZ Biolabs employs high-resolution mass spectrometry platforms (e.g., Thermo Fisher Orbitrap Fusion Lumos) alongside specific antibody-based crotonylated peptide enrichment strategies to enhance the sensitivity of detecting low-abundance crotonylation modifications. Additionally, advanced data acquisition techniques such as Data-Independent Acquisition (DIA) and Data-Dependent Acquisition (DDA) enable accurate site-specific identification and quantification.
Analysis Workflow
MtoZ Biolabs has developed a highly efficient crotonylation proteomics analysis workflow that covers every step from sample preparation to bioinformatics interpretation, ensuring high reproducibility and in-depth coverage.
1. Protein Extraction: Extraction of total proteins from cells, tissues, or biofluids while maintaining protein integrity.
2. Enzymatic Digestion: Proteins are enzymatically digested using trypsin or Lys-C to generate peptides suitable for mass spectrometry analysis.
3. Crotonylated Peptide Enrichment: Specific anti-crotonylation antibodies are used to enrich modified peptides, increasing detection sensitivity and specificity.
4. Peptide Labeling (TMT/iTRAQ or Label-Free):
1. TMT/iTRAQ: Ideal for high-throughput quantitative comparisons across multiple samples.
2. Label-Free: Enables large-scale protein identification and quantification without isotope labeling.
5. LC-MS/MS Detection: High-resolution mass spectrometry platforms such as Orbitrap or Q Exactive HF are employed to precisely identify crotonylation modification sites.
6. Data Analysis: Advanced bioinformatics tools, including MaxQuant and Proteome Discoverer, are used for peptide spectrum matching, modification site identification, and quantitative analysis. Functional insights are further explored through Gene Ontology (GO) and KEGG pathway enrichment analysis.
Service Advantages
✅ Ultra-Sensitive Detection: Specific antibody-based enrichment combined with Orbitrap high-resolution mass spectrometry enhances the detection of low-abundance crotonylation modifications.
✅ High-Precision Quantification: Supports both TMT/iTRAQ-based labeled quantification and label-free quantification, catering to different experimental needs.
✅ Comprehensive Data Interpretation: Provides crotonylation site identification, protein function annotation, and pathway enrichment analysis to facilitate biological insights.
✅ Broad Sample Compatibility: Suitable for various biological samples, including cells, tissues, serum, plasma, and urine.
Sample Submission Suggestions
To ensure high-quality crotonylation proteomics analysis data, MtoZ Biolabs has specific requirements for sample type, quality, and storage conditions. The following sample types are commonly accepted:
Proper sample handling, storage, and transportation are critical to ensuring optimal experimental results. MtoZ Biolabs provides a detailed sample submission guide to ensure the best analytical outcomes. For specialized sample types or custom preparation needs, please contact our technical support team to optimize your experimental workflow.
Applications
Crotonylation proteomics analysis has become an emerging focus in various biological and medical fields. MtoZ Biolabs' Crotonylation Proteomics Analysis Service supports research in the following areas:
Case Study
Case 1
A study using high-resolution LC-MS/MS systematically identified crotonylated proteins in human embryonic stem cells (hESCs). The research focused on the glycolytic enzyme GAPDH, demonstrating how crotonylation regulates its activity and metabolic pathways during hESC differentiation. Through in-depth proteomic analysis, the study provided mechanistic insights into crotonylation’s role in cellular metabolism and differentiation.
Zhang, J. et al. Stem Cell Res. Ther. 2023.
By leveraging the Crotonylation Proteomics Analysis Service, we can provide precise mass spectrometry-based analysis to detect and quantify crotonylation sites, advancing research on stem cell differentiation and metabolic regulation.
MtoZ Biolabs utilizes state-of-the-art mass spectrometry platforms to deliver high-quality Crotonylation Proteomics Analysis Service. For service inquiries or technical support, please contact our expert team today!
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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