The proteolytic process is an irreversible post-translational modification (PTM) that regulates many cellular processes, including gene expression, embryogenesis, cell cycle, programmed cell death, intracellular protein targeting, and endocrine/neural function. Creative BioMart's extensive experience in post-translational modification of proteins based on computational and experimental methods enables us to provide protein proteolytic service to accelerate your project.

Proteolysis refers to the limited and highly specific hydrolysis of peptide and isopeptide bonds of proteins by a special class of enzymes called proteases. Proteolytic products form new N and C termini. Proteolysis can lead to activation, inactivation, and complete alteration of protein function, and they regulate a wide variety of biological processes, including DNA replication, cell cycle progression, cell proliferation, and cell death, as well as in autoimmune diseases, bacterial infections, cancer, and viral disease and other human diseases. Proteases make extensive, irreversible post-translational modifications to the structure and biological function of proteins. More than 30 drugs targeting these enzymes are currently approved for clinical use. Fully understanding the cellular function of a given protease holds promise for new insights into new proteolytic pathways in vivo and new drug targets for disease.

Fig 1. Schematic overview of degradomics approaches using positive selection of N-terminal peptides. (Klein T, et al., 2020)

Services

To fully understand proteolytic post-translational modifications, it is essential to understand the proteins processed by proteases, as well as the functions and specific processing events of these substrates. Degradomics is currently the application of high-throughput methods to study proteases, their substrates, and their inhibitors on a system-wide scale. Creative BioMart has established a professional protein proteolytic post-translational modification service platform, developed a variety of proteomic-based methods to identify protease substrates and cleavage sites, and characterize protein N- and C-termini. These methods are able to identify thousands of ends in vitro and in vivo and in complex tissues. In addition, our scientists are commmitted to developing software tools for generating quantitative confidence and isoform assignment scores as well as automatic annotation of N-terminal peptides to determine their position in the protein sequence. We provide the following services for protein hydrolysis, including but not limited to:

• Dentifying protease active site specificity: we used an array and library-based approach to identify protease active site specificity. We can help you elucidate the cleavage site specificity of many proteases from all catalytic classes and also reliably identify relevant in vivo protease substrates from the cleavage site.
• Identification of protease substrates: we used a proteomic approach to identify protease substrates, aimed at identifying substrates from complex proteomes and identifying multiple cleavage sites in proteins in vitro.
• Characterizing protein N- and C-termini: N-terminal or C-terminal post-translational modifications, including proteolytic processing, can greatly affect the localization and activity of many proteins. We have developed various telopeptide enrichment strategies to improve the coverage and dynamic range of telopeptide identification.

Approaches for Analyzing Protein Proteolytic

• Peptide libraries and microarrays.
• Two-dimensional polyacrylamide gel electrophoresis (PAGE) technique.
• Based on LC-MS/MS technology.
• Substrate Terminal Amine Isotope Labeling (TAILS).
• C-terminal amine-based substrate isotope labeling (C-TAILS).
• Combined strong cation exchange chromatography.
• High-throughput database.

Our mission is to provide global customers with the most comprehensive and professional customized protein proteolytic service. If you have any special requirements about our services, please feel free to contact us. We are looking forward to working together with your attractive projects.

References

1. Klein T, Eckhard U, Dufour A, et al.. (2018) Proteolytic Cleavage-Mechanisms, Function, and "Omic" Approaches for a Near-Ubiquitous Posttranslational Modification. Chem Rev. 118(3): 1137-1168.
2. Rogers LD, Overall CM. (2013) Proteolytic post-translational modification of proteins: proteomic tools and methodology. Mol Cell Proteomics. 12(12): 3532-3542.

• Protein Acetylation Service
• Protein Carbonylation Service
• Protein Glycosylation Service
• Protein Hydroxylation Service
• Protein Methylation Service
• Protein Nitrosylation Service
• Protein Phosphorylation Service
• Protein Sulfation Service
• Protein Ubiquitination Service