Lipidation Analysis Service

Lipidation is a crucial post-translational modification (PTM) involving the covalent attachment of lipid moieties (e.g., myristoylation, palmitoylation, prenylation, and glycosylphosphatidylinositol (GPI) anchoring) to specific amino acid residues, regulating membrane localization, signal transduction, protein stability, and cellular interactions. Dysregulated lipidation is implicated in cancer, neurodegenerative disorders, metabolic diseases, and infections. For example, aberrant palmitoylation contributes to oncogenesis, while defective prenylation disrupts Ras-mediated signaling pathways. Targeting lipid-modifying enzymes, such as acyltransferases (e.g., NMT, DHHC), lipid hydrolases (e.g., APT), and lipidation reader proteins, presents novel therapeutic opportunities. Small-molecule inhibitors targeting lipidation-related enzymes are under development for cancer and neurological disorders.   

 



Figure 1. The Most Common Type of Lipidation

 

Services at MtoZ Biolabs  

leverages cutting-edge mass spectrometry and bioinformatics tools, MtoZ Biolabs offers lipidation analysis service to provide site-specific lipidation profiling, facilitating the discovery of novel biomarkers, understanding disease mechanisms, and identifying potential therapeutic targets associated with aberrant lipidation. Our lipidation analysis service includes:  

✔ Global lipidation site identification (LC-MS/MS-based profiling)  

✔ Quantitative lipidation dynamics (TMT/iTRAQ labeling strategies)  

✔ Functional validation (site-directed mutagenesis and activity assays)  

✔ Customized panels for specific lipid types (myristoyl, palmitoyl, prenyl, GPI, etc.)  

✔ Bioinformatics analysis (lipidation-associated pathways and disease correlations)  

 

Sample Compatibility: Cells, tissues, serum, exosomes, and other complex biological samples.  

 

Analysis Workflow  

1. Sample Preparation: Protein extraction, enzymatic digestion, lipidated peptide enrichment (click chemistry, affinity purification).  

2. LC-MS/MS Analysis: High-resolution Orbitrap platforms for deep lipidation coverage.  

3. Data Processing: MaxQuant/Percolator for lipidation site identification.  

4. Bioinformatics: Metascape/KEGG analysis for lipidation-related pathways.  

5. Report Generation: Detailed summary with validation-ready data.  

 



Figure 2. The Workflow of Lipidation Analysis

 

Service Advantages  

✅ Ultra-High Sensitivity: Detects low-abundance lipidated peptides (≥0.1 fmol).  

✅ Comprehensive Coverage: Multi-lipid type analysis within a single workflow.  

✅ Speed & Precision: 2-week turnaround with >95% site identification accuracy.  

✅ Expert Support: Tailored study design and data interpretation by lipidation specialists.  

 

Case Study

1. Chemical Proteomics Strategies for Analyzing Protein Lipidation Reveal the Bacterial O-Mycoloylome

Protein O-mycoloylation is a unique lipidation modification in the outer membrane of Corynebacterium species, where hydrophobic mycolic acids are attached to serine residues to regulate membrane protein localization and function. This study established a chemical proteomics strategy combining metabolic labeling, click chemistry, cleavable linkers, and novel LC-MS/MS methods to overcome enrichment and detection challenges in analyzing lipidated peptides. In Corynebacterium glutamicum, approximately 30 candidate O-mycoloylated proteins—including porins, mycoloyltransferases, and secreted hydrolases—were identified, most associated with cell envelope functions and exhibiting spatially clustered modification sites. Site-specific analysis revealed a preference for serine residues within conformationally flexible peptide motifs. This work provides the first global profile of the O-mycoloylome, offers insights into the mycomembrane proteome of pathogens (e.g., Mycobacterium tuberculosis), and presents broadly applicable chemical approaches for studying protein lipidation. Our lipidation analysis service specializes in comprehensive analysis of protein lipidation, leveraging innovative chemical proteomic strategies to identify and characterize lipid-modified proteins in complex biological systems. Optimized for hydrophobic or mass spectrometry-sensitive modifications (e.g., acylation, prenylation), the platform supports targeted studies of membrane-associated proteins in bacterial and eukaryotic systems.

 



Banahene, N. et al. J Am Chem Soc. 2024. 

Figure 3. Identification of Modified Peptides Using Cleavable Linker and LC-MS/MS Strategies

 

2. Liquid-liquid extraction of lipidated peptides for direct identification of lipidation sites

Proteins undergo lipidation through various modifications, including myristoylation, palmitoylation, farnesylation, and geranylgeranylation. Traditional chemical proteomics methods can identify lipidated proteins but face limitations in detecting endogenous lipidation sites and in vivo applicability. This study introduces a liquid-liquid extraction strategy combined with LC-MS/MS using a high-concentration acetonitrile gradient to directly enrich and analyze hydrophobic lipidated peptides. In HeLa cells, 90 lipidation sites were identified, including 75 N-terminal myristoylation sites, along with dually modified peptides. Application in mouse tissues further revealed tissue-specific lipidation profiles, providing a powerful tool for the systematic analysis of protein lipidation. The Lipidation Analysis Service integrates an optimized lipidated peptide separation strategy with high-sensitivity mass spectrometry for precise identification of lipidation sites. By adapting different enzymatic digestion strategies, this service extends coverage of lipidation types and enables the identification of dually modified peptides and tissue-specific lipidation patterns. It is designed for endogenous protein lipidation analysis, offering a tailored solution for detecting lipid modifications in complex biological systems.

 



Kazuya, Tsumagari. et al. bioRxiv. 2023.

Figure 4. Lipidation Profiling of HeLa Cell Proteins

 

Applications  

1. Basic Research: Unraveling lipidation’s role in signaling and metabolism.  

2. Biomarker Discovery: Identifying lipid-based disease markers.  

3. Drug Development: Validating lipidation-modifying enzymes as therapeutic targets.  

4. Precision Medicine: Stratifying patients based on lipidation profiles.

 

How Can We Help You?  

1. Communication of Requirements: Fill out an inquiry form for a customized solution.  

2. Sample Submission: We accept dry ice shipments and samples such as cells (1×10⁷), body fluids (>200 µL), and tissues (>50 mg). Contact us for specific requirements.  

3. Experiment and Analysis: Tailored lipidation analysis is completed within 15 business days, with real-time project updates for complex studies.  

4. Report Delivery: Reports include lipidation site identification, quantitative modification data, raw MS files, peptide spectra (MS/MS), and bioinformatics analysis results.  

 

Lipidation is fundamental to cellular function and disease progression. MtoZ Biolabs integrates advanced technologies with deep PTM expertise to deliver high-quality lipidation analysis service, bridging fundamental research and translational applications. Partner with us to decode lipidation modifications and unlock new therapeutic possibilities.