Carbamylation Proteomics Service

Post-translational modifications (PTMs) are essential regulatory mechanisms that shape protein structure and function, with broad impact on cell fate, signal transduction, and metabolic balance. Among them, carbamylation is a non-enzymatic PTM induced by isocyanic acid, a byproduct of urea metabolism. This modification mainly targets lysine residues, resulting in the formation of homocitrulline. Protein carbamylation is widely present in pathological contexts such as chronic kidney disease (CKD), inflammation, atherosclerosis, and autoimmune disorders, and has emerged as a promising functional modification and biomarker.

 



Zhong, Q. et al. MedComm (2020). 2023.

Figure 1. The Different Pathways of Carbamylation And Its Roles in Several Diseases

 

However, the non-specific nature of carbamylation and its close mass shift to other modifications such as acetylation present challenges in its detection and quantification via conventional proteomic approaches. To better understand the biological relevance of carbamylation, a high-specificity, high-throughput analytical strategy is urgently needed.

 

Leveraging years of experience in proteomics, MtoZ Biolabs has developed a dedicated high-resolution mass spectrometry platform tailored for Carbamylation Proteomics Service. Combined with targeted enrichment methods and advanced bioinformatics analysis, our Carbamylation Proteomics Service provides researchers with accurate, efficient, and interpretable results—accelerating the exploration of carbamylation-mediated regulation in both physiological and pathological contexts.

 

Analysis Workflow

1. Sample Preparation

• Compatible with various sample types, including tissues, cells, serum, and plasma
• Optimized to avoid urea-induced artificial carbamylation during preparation

 

2. Protein Digestion

Enzymatic digestion using trypsin or other standard proteases

 

3. Enrichment of Modified Peptides

Affinity enrichment using anti-acetyllysine antibodies with cross-reactivity to carbamylated peptides

 

4. Mass Spectrometry Analysis

High-resolution LC-MS/MS analysis using Orbitrap or equivalent systems

 

5. Data Processing and Annotation

• Site identification via software such as Mascot and MaxQuant
• Functional analysis based on GO, KEGG, and other databases

 

6. Deliverables

• Lists of identified proteins and modification sites
• Enrichment plots and pathway analyses
• Raw spectra, quantitative matrices, and a comprehensive project report

 

Service Advantages

1. High-Throughput Detection: Covers thousands of modified peptides and hundreds of proteins—suitable for both screening and mechanistic studies

2. Specific Enrichment Strategy: Combines antibody-based enrichment with high-resolution MS to distinguish carbamylation from other PTMs

3. Sample Versatility: Supports various biological matrices for both basic research and clinical studies

4. PTM Crosstalk Insights: Expandable to multi-PTM studies such as acetylation, phosphorylation, and methylation

5. Standardized Quality Control: Built-in technical replicates and QC checkpoints throughout the workflow

6. Integrated Bioinformatics Support: Provides functional interpretation and pathway visualization to support downstream research

 

Applications

1. Chronic Kidney Disease Research

Investigate the effects of high-urea environments on protein carbamylation and renal injury mechanisms

 

2. Cardiovascular Disease Mechanisms

Explore the involvement of carbamylated proteins in the development of atherosclerosis and related disorders

 

3. Inflammation and Immune Modulation

Analyze the regulatory role of carbamylation in immune cells such as macrophages

 

4. Aging-Related Protein Regulation

Assess age-dependent variation in carbamylation levels and tissue specificity

 

5. Biomarker Discovery

Identify disease-associated carbamylated proteins as potential diagnostic or prognostic markers

 

6. Drug Mechanism Studies

Monitor the regulatory effects of therapeutic interventions on carbamylation dynamics

 

Case Study

1. Improvement of Shotgun Proteomics in the Negative Mode by Carbamylation of Peptides and Ultraviolet Photodissociation Mass Spectrometry

This study addresses the challenge of poor ionization efficiency of acidic peptides in positive-mode electrospray ionization (ESI) during bottom-up proteomics. The authors introduce a derivatization strategy based on carbamylation, which converts lysine residues and N-terminal amines into less basic amides, thereby improving ionization efficiency in negative mode. Combined with ultraviolet photodissociation mass spectrometry (UVPD-MS), this approach was applied to tryptic peptides from Halobacterium salinarum. LC/UVPD-MS analysis of carbamylated samples resulted in a 45% increase in peptide identifications and a 25% increase in protein identifications compared to unmodified digests under the same negative mode conditions. The findings demonstrate a significant improvement in shotgun proteomics performance in the negative ionization mode. Carbamylation Proteomics Service leverages peptide carbamylation combined with high-resolution mass spectrometry to enhance the detection of acidic proteins or peptides under negative ion mode. This approach improves proteome coverage and is well-suited for comprehensive profiling of standard and extremophile-derived samples. The workflow is compatible with high-throughput LC-MS/MS analysis.

 



Greer, SM. et al. Anal Chem. 2014.

Figure 2. Shotgun Proteomics Analysis of Carbamylation of Peptides

 

2. Analysis of a Macrophage Carbamylated Proteome Reveals a Function in Post-Translational Modifcation Crosstalk

This study systematically profiled the carbamylated proteome of LPS-stimulated macrophages using an affinity strategy based on the cross-reactivity of anti-acetyllysine antibodies. A total of 8,923 carbamylated peptides were identified. Integrated analysis with acetylation and phosphorylation datasets revealed 1,183 proteins modified by all three PTMs. Notably, carbamylation was found to interfere with the activity of the deubiquitinase OTULIN within the ubiquitin–proteasome pathway, suggesting that carbamylation may play a regulatory role in PTM crosstalk and immune-related signaling networks. Carbamylation Proteomics service offers integrated enrichment and mass spectrometry-based analysis to systematically characterize carbamylation profiles under various cellular conditions. It enables the investigation of crosstalk between carbamylation and other post-translational modifications, supporting studies on inflammatory models, immune regulation, and protein function. This service is suitable for uncovering the broader regulatory landscape of protein carbamylation.

 



You, Y. et al. Cell Commun Signal. 2023.

Figure 3. Carbamylated Proteomics Analysis of Macrophages Treated with Bacterial Lipopolysaccharide

 

FAQ

Q1: Is carbamylation a reversible modification?

A1: No. Carbamylation is an irreversible, non-enzymatic modification typically induced by isocyanic acid and cannot be enzymatically removed.

 

Q2: Can other PTMs be analyzed simultaneously?

A2: Yes. We support multiplex PTM analysis, including phosphorylation, acetylation, and methylation.

 

Q3: Will residual urea in the sample interfere with results?

A3: Yes. High levels of urea may introduce artificial carbamylation. We recommend avoiding strong denaturants during sample preparation.

 

Q4: Is quantitative analysis available?

A4: Yes. Both label-free quantification and TMT/iTRAQ labeling strategies are supported.

 

Q5: What is the turnaround time for the service?

A5: Typically 3–6 weeks, depending on sample number and project complexity.

 

MtoZ Biolabs is committed to delivering standardized and reliable proteomics solutions. Our Carbamylation Proteomics Service is ideal for basic research, disease mechanism studies, and biomarker discovery. Contact us today for more details or technical consultation.