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Veterinary Therapeutic Protein and Peptide Post-Translational Modification Analysis
Post-translational modifications (PTMs) change the characteristics of proteins and peptides through hydrolytic cleavage and the addition of modifying groups to specific amino acids, affecting normal cellular functions and disease pathways. BioVenic provides specialized and comprehensive analysis services for PTMs in veterinary therapeutic proteins and peptides. Utilizing cutting-edge mass spectrometry and bioinformatics methods, we deliver detailed identification and analysis of chemical modifications occurring after biosynthesis. This service aids our clients in progressing their preclinical research on veterinary therapeutic proteins and peptides.
Protein and Peptide PTMs Overview
PTMs are critical chemical changes that proteins and peptides undergo after their synthesis, affecting their function, stability, and interactions. BioVenic provides specialized therapeutic protein and peptide PTM analysis services for veterinary medicine preclinical research. These modifications encompass various modifications and our advanced analytical techniques help elucidate these modifications on proteins and peptides, offering insights essential for veterinary therapeutic development.
Fig.1 Overview of common PTMs in proteins. 1, 2
Protein and Peptide PTM Analysis Services
Our PTM analysis services encompass a broad range of modifications, extending beyond the commonly studied ones like phosphorylation and ubiquitination. We offer comprehensive analysis to identify and characterize a wide array of PTMs, providing detailed insights into the molecular mechanisms governing protein function. The following table provides an overview of the types of modifications that BioVenic specializes in analyzing.
| PTMs | Functions |
|---|---|
| Oxidation | Affects protein structure, catalytic function, and signaling processes, commonly seen in studies related to oxidative stress and aging. |
| Deamidation | Causes reversal, extension, and aging of proteins and peptides. |
| Glycosylation | Influences protein stability, activity, and intercellular interactions, involved in immune responses, cell recognition, and tumor development. |
| Phosphorylation | Plays a key role in signal transduction, cell cycle regulation, and metabolic regulation. |
| Methylation | Affects DNA, histones, and non-histone proteins, involved in epigenetic regulation and signal transduction. |
| Ubiquitination | This leads to protein degradation, affecting protein stability regulation, cell cycle, and DNA repair. |
| Acetylation | Regulates cellular physiological metabolism, such as DNA transcription, mRNA splicing, and apoptosis. |
| Lipidation | Affects protein-membrane binding and cellular localization, involved in cell signal transduction and membrane protein function. |
| Hydroxylation | Stabilizes protein structure, regulates activity, and influences cellular interaction. |
| SUMOylation | Modulates protein function, protects against degradation, regulates transcription, and affects DNA repair. |
Mass Spectrometry-Based Analysis
Mass Spectrometry is one of the most widely applied analytical techniques in the study of PTMs. It determines protein modifications, including the type of modification, its location, and relative abundance, thereby offering deep insights into the function and regulatory mechanisms of proteins.
Fig.2 PTM Workflow Schematic: Overview of key steps in sample prep, PTM enrichment, MS, and bioinformatics. 3, 4
- PTM-Specific Enrichment
Enrichment specific to PTMs is an essential preparatory step before analyzing protein samples with mass spectrometry. PTMs are frequently found in low abundance within veterinary therapeutic proteins and peptides, and their presence can be fleeting and unstable. This makes the direct detection of PTMs in complex biological samples particularly challenging. To address these issues, BioVenic has developed a platform incorporating specific enrichment techniques, employing various methods to improve detection sensitivity and accuracy.
| Services | Description |
|---|---|
| Ion Exchange Chromatography | By adjusting the pH and ionic strength, selectively bind and elute specific modified proteins or peptides. |
| Immobilized Metal Ion Affinity Chromatography | Effectively enrich phosphorylated or other metal-affinity modified proteins or peptides through binding with metal ions such as iron, nickel, or zinc. |
| Immunoaffinity Chromatography | Use specific antibodies to capture proteins or peptides with particular PTMs. |
- Mass Spectrometry
BioVenic uses a "bottom-up" proteomics approach for PTM analysis, breaking down veterinary therapeutic proteins and peptides into smaller fragments using specific proteases, typically trypsin, before conducting mass spectrometry (MS) analysis. Our lab is equipped with high-sensitivity, high-resolution instruments, such as Fourier transform (FT) mass spectrometers.
Bioinformatics Analysis
Managing and interpreting large datasets from LC-MS/MS is crucial, especially for identifying PTMs and validating experimental results. BioVenic employs advanced bioinformatics tools and algorithms for comprehensive analysis of LC-MS/MS data, providing precise information on post-translational modifications in veterinary therapeutic proteins and peptides.
PTM Prediction Algorithms
Our experts use algorithms like Ascore, PTMScore, PhosphoRS, and Mascot Delta Score to predict specific PTMs from LC-MS/MS data. These tools help assess the likelihood of modification sites, aiding researchers in identifying which modifications are present.
Validation of Modification Sites
We validate modification sites identified in LC-MS/MS by comparing experimental data with theoretical predictions, such as expected mass spectra, to ensure accuracy and reliability.
Service Workflow
Our service for veterinary therapeutic proteins and peptides begins with an initial consultation to understand the client's needs and project scope. This is followed by PTM-specific enrichment to enhance post-translational modifications for precise analysis. Mass spectrometry is then used to accurately identify and quantify proteins and peptides. The data is processed using advanced bioinformatics tools and prediction algorithms to provide insights into interactions and functions. Finally, the results are validated, and a comprehensive report is delivered, detailing the findings and their implications.
Fig.3 The veterinary therapeutic protein and peptide PTM determination service workflow. (BioVenic Original)
Why Choose Us?
Equipped with state-of-the-art mass spectrometry equipment and bioinformatics tools to provide high-precision and high-resolution analysis results.
Not limited to common PTM types such as phosphorylation, acetylation, and ubiquitination, but also more complex and rare modification types.
Our team has extensive bioinformatics and proteomics expertise and utilizes advanced algorithms and software tools to predict and validate PTM sites.
PTMs significantly impact the function, stability, and biological activity of veterinary therapeutic proteins and peptides. BioVenic offers PTM analysis services for veterinary therapeutic proteins and peptides, accurately identifying and quantifying various PTMs, and providing crucial molecular-level information for the development of veterinary therapeutic drugs. If you are interested, please contact us now.
References
- Salas-Lloret, Daniel, and Román González-Prieto. "Insights in post-translational modifications: ubiquitin and SUMO." International Journal of Molecular Sciences 23.6 (2022): 3281.
- Image retrieved from Figure 1 "Post-translational modifications of proteins". Salas-Lloret, Daniel, and Román González-Prieto. 2022, used under CC BY 4.0, the title was changed to " Overview of common PTMs in proteins".
- Pascovici, Dana, et al. "Clinically relevant post-translational modification analyses—maturing workflows and bioinformatics tools." International journal of molecular sciences 20.1 (2018): 16.
- Image retrieved from Figure 1 "Schematic PTM Workflow illustrating some of the common steps associated with sample preparation, PTM enrichment, MS and bioinformatics analysis". Pascovici, Dana, et al., 2018, used under CC BY 4.0, the title was changed to " "PTM Workflow Schematic: Overview of key steps in sample prep, PTM enrichment, MS, and bioinformatics".