Creative BioMart is a well-known expert who committed to developing a variety of methods to explore enzyme sequences and create new high-efficiency biocatalysts. With years of experience, we provide customized protease substrate screening service to precisely meet customer requirements.

Introduction of Protease Substrate

Proteases are modulators of many biological pathways, including protein catabolism, antigen processing and inflammation, as well as various disease conditions such as malignant metastasis, viral infection and parasite invasion. Identifying the substrate of a given protease is critical to understanding its function, and this information can also help design specific inhibitors and active site probes. Considering that the primary structure of all known proteases will soon be available, an important challenge is to determine the structure-activity relationship that governs the hydrolysis of the substrate. To address this challenge in protease research, a range of methods have been developed to identify natural protein substrates as well as map the overall substrate specificity patterns of proteases. The combined method of substrate phage display has been proved to be effective in providing a rapid and systematic method to determine the substrate specificity of a variety of proteases, and it is possible to develop new protease-specific tools and therapeutics.

Fig 1. Schematic illustration of a phage-based approach to discover protease substrate peptides. (Chen S, et al., 2020)

Services

Traditional high-throughput proteomics methods are used to identify and prioritize enzyme substrates. However, substrate screening experiments for new enzymes are laborious, time-consuming and costly. As a leading protein engineering service provider, Creative BioMart has established a powerful platform for screening the substrate specificity of proteases to achieve protein turnover and functional activation of substrate proteins. We have developed a method to use the phage library to discover the substrate protease, in which more than 10⁷ potential substrates can be tested concurrently. It can help you to quickly investigate the substrate recognition and specificity of individual proteases or closely related members.

Our scientists are committed to constructing a fusion library containing random substrate sequences located between the binding domain of filamentous bacteriophage M13 and gene Ⅲ coat protein. The coat protein displays the fusion protein and encapsulates the gene it encodes in internal. Substrate phage screening has shown great practical value in quickly determining the specificity of proteases and characterizing the substrate recognition spectrum of proteases, and is widely used by customers around the world. Compared with other methods, substrate phage display screening has the following advantages:

1. The construction of a substrate phage library does not require expensive reagents or equipment. The system is suitable for laboratories of all sizes.
2. The substrate phage display screening is fast and very robust. The entire screening process, including solution screening and solid phase screening, can be completed within three months without the need to modify the screening scheme.
3. The screening procedure is very flexible and can be simplified into a high-throughput format easily.
4. Compared with synthetic combinatorial peptide screening libraries, the phage system is the ability to obtain accurate but not average substrate/protease interaction data. The synthetic screening method can only get the average result for each position.

In addition, we try to identify proteases by designing and constructing substrate libraries based on small molecule fragments, synthetic and biological methods to generate peptide libraries to discover protease substrates, and develop chemical tools for protease detection or inhibition. We will work with you to develop the most appropriate strategy and provide the most meaningful data for your research for accelerating the research of life sciences. If you are interested in our services, please do not hesitate to contact us for more information.

References

1. Chen S, Yim J J, Bogyo M. (2020) Synthetic and biological approaches to map substrate specificities of proteases. Biological chemistry. 401(1): 165-182.
2. Sedlacek R, Chen E. (2005) Screening for protease substrate by polyvalent phage display. Comb Chem High Throughput Screen. 8(2): 197-203.

• Enzyme Activity Analysis Service
• Enzyme Stability Enhancement Service
• Protease Substrate Specificity Profiling Service