Disulfide bonds are a common posttranslational modification in proteins, which are formed between the sulfur atoms of two cysteine (Cys) residues during the biosynthesis of proteins. Due to its wide presence, analysis of disulfide bonds is often critical to enzyme discovery and identification.

Creative Enzymes is an industry leader in the determination and analysis of enzyme structures, providing comprehensive and effective solutions for enzyme disulfide bond analysis based on our advanced technology platforms and assays.

The Role of Disulfide Bonds on Enzymes

Compared to other proteins, disulfide bonds are especially critical for enzymes in proper folding, and they have both structural and functional roles. For example, non-native disulfide bonds can be engineered into enzymes to increase their stability. Also, some disulfide bonds, known as allosteric disulfides, are responsible for the effective biological functions of enzymes, and the cleavage of such bonds would lead to a change in the protein activity. Therefore, disulfide bond analysis provides key information to research pertaining to protein stability, structure-function relationships, and any disulfide-mediated isoforms of enzymes.

Significance of Enzyme Disulfide Bond Analysis

Disulfide bond assays provide researchers with important tools and means to gain insight into enzyme structure and function. We are committed to disulfide bond analysis, and we continue to utilize a variety of state-of-the-art techniques to promote the progress of disulfide bond research. By revealing the properties and mechanisms of disulfide bonds, we can help researchers better understand the biological functions of enzymes and provide new directions and strategies for the study of related diseases and drug development.

Analytical Techniques for Disulfide Bonds

Disulfide bonds can be analyzed mainly by three classes of techniques as follows.

• Crystallographic techniques producing high-resolution three-dimensional structures.
• Algorithmic techniques based on sequence data.
• Techniques combining experimental and algorithmic methods to analyze a mixture of digested peptides.

A variety of analytical techniques are involved in a typical study, such as NMR spectroscopy, X-ray crystallography, Edman degradation, diagonal paper electrophoresis, and liquid chromatography (LC) coupled with mass spectrometry (MS). Compared to limitations in other methods, such as high purity sample requirements and low throughput, bottom-up MS is the most widely used method for disulfide bond analysis. In general, this bottom-up approach involves the digestion of large biomolecules into small pieces that are easy to analyze, with the help of ionization techniques, complementary fragmentation techniques, and an LC system for separation.

Figure 1 A typical disulfide bond (DSB) analysis workflow.
(Analytical and Bioanalytical Chemistry 2018)

Creative Enzymes provides a highly sensitive HPLC-MS/MS platform that can analyze disulfide bonds in multiple samples from various sources. In addition, we develop advanced machine-learning strategies based on sequence-based information. Furthermore, we provide services regarding disulfide artifacts verification and disulfide engineering. Our featured services include as following.

• Algorithmic analysis based on sequence data.
• Disulfide bond analysis by HPLC-MS/MS platform.
• Disulfide artifacts verification.
• Disulfide engineering.

If you are interested in us, please feel free to contact us.