γ-Carboxylation Proteomics Service

γ-Carboxylation proteomics is a specialized, high-resolution approach designed to systematically characterize γ-carboxyglutamic acid (Gla) modifications and their effects on protein conformation, calcium-binding affinity, and functional activity. Leveraging cutting-edge mass spectrometry and Gla-targeted enrichment strategies, this workflow enables precise detection and localization of Gla-modified peptides, providing vital molecular insights into coagulation, bone metabolism, and vascular calcification pathways.

 

γ-Carboxylation is a vitamin K–dependent post-translational modification catalyzed by γ-glutamyl carboxylase in the endoplasmic reticulum. It converts glutamate residues into Gla, endowing proteins with calcium-binding capacity—essential for the activity of coagulation factors (e.g., FII, VII, IX, X), osteocalcin, and matrix Gla protein (MGP). Deficiencies in Gla modification are implicated in bleeding disorders, osteoporosis, and arterial calcification.

 



Source: Wikipedia

Figure 1. Vitamin K-Dependent γ-Carboxylation Mechanism

 

MtoZ Biolabs provides γ-Carboxylation Proteomics Service that supports reliable profiling of Gla modifications, functional annotation and biomarker discovery in both basic and translational research using a high-resolution mass spectrometry platform.

 

Services at MtoZ Biolabs

Our γ-Carboxylation Proteomics Service integrates multiple MS and enrichment strategies to elucidate the relationship between modification sites and structural-functional regions:

·  Gla-specific peptide enrichment: Enhanced recovery of Gla-modified peptides using calcium-affinity techniques or Gla-specific antibodies

·  Site localization via tandem MS/MS: Confident site-level annotation using optimized HCD-based fragmentation and PTM-aware database searches

·  Quantitative modification profiling: Differential analysis of Gla abundance across biological states or treatment groups

·  Mapping of multi-site modification heterogeneity: Explore Gla modification patterns and co-occurrence across protein domains

·  Publication-ready reporting: Includes annotated site lists, peptide spectra, quantitative charts, and enrichment analysis results

 

Why Choose MtoZ Biolabs?

1. Advanced Analysis Platform: MtoZ Biolabs established an advanced γ-Carboxylation proteomics analysis platform, guaranteeing reliable, fast, and highly accurate analysis service.

2. Ultra-Sensitive Gla Enrichment Workflow: Designed to detect low-abundance Gla peptides with high specificity and minimal background.

3. Integrated Cross-Modification Analysis: Enables multi-omics integration with phosphorylation, glycosylation, and expression datasets.

4. High-Data-Quality: Deep data coverage with strict data quality control. AI-powered bioinformatics platform integrate all proteomics analysis data, providing clients with a comprehensive data report.

5. One-Time-Charge: Our pricing is transparent, no hidden fees or additional costs.

 

Applications

The γ-Carboxylation Proteomics Service can be broadly applied to explore the roles of Gla modifications in physiological and pathological regulation, including:

·  Coagulation factor activation analysis
Map site-specific Gla modifications in vitamin K–dependent clotting proteins (FII, VII, IX, X)

·  Vitamin K response assessment
Monitor how dietary deficiency or drug intervention affects carboxylation efficiency

·  Bone metabolism studies
Correlate osteocalcin Gla status with bone density, turnover rate, or fracture susceptibility

·  Cardiovascular and vascular calcification research
Evaluate MGP carboxylation levels as predictive markers for arterial stiffness or calcification

 

Case Study

Case1：Quantitative Profiling of Prothrombin γ-Carboxylation Using LC–MS/MS

In a 2023 study, researchers developed an advanced LC–MS/MS protocol for Gla proteoform analysis in human plasma. By applying alkaline mobile phases and polarity switching, the method addressed CO₂ loss and fragmentation issues that previously hampered Gla peptide detection. From as little as 10 μL of plasma, multiple prothrombin Gla forms were successfully quantified.

 

Comparative analysis between healthy donors and warfarin-treated patients revealed a significant elevation of under-carboxylated prothrombin precursors in the latter group, demonstrating the direct impact of Gla deficiency on bioactivity. Non-Gla peptides served as complementary quantifiers, enabling absolute quantification of vitamin K–dependent proteins using TPA methodology.

 



Singh, D. K. et al. Anal Chim Acta. 2023.

 

Collectively, these findings underscore the robustness of γ-Carboxylation Proteomics Service in clinical sample analysis, isomer quantification, and PTM site validation.

 

FAQ

Q1: What makes γ-carboxylation site detection particularly challenging in MS-based proteomics?

Gla-modified peptides are prone to decarboxylation during digestion or ionization, and mass shifts are subtle. This can result in signal suppression or under-reporting of sites. MtoZ Biolabs applies protective digestion protocols, Gla-specific enrichment, and high-energy fragmentation combined with tailored database parameters to ensure confident detection.

 

Q2: Can Gla modification be analyzed alongside vitamin K status?

Yes. We can integrate Gla proteomics data with LC–MS-based vitamin K quantification to assess enzyme activity, modification efficiency, and metabolic status—ideal for drug response studies or nutritional intervention models.

 

Q3: Do you offer orthogonal validation methods?

Absolutely. We support optional validation using Western blot, ELISA, or antibody-based enrichment. Joint annotation with glycosylation or phosphorylation datasets is also available to enhance biological interpretation.

 

MtoZ Biolabs’ γ-Carboxylation Proteomics Service delivers a full-spectrum solution for vitamin K–dependent modification studies—from site-specific annotation to pathway modeling. Our customizable workflows support mechanistic research, therapeutic evaluation, and biomarker discovery. Contact us to learn more about project customization.