Biotin Detection of Protein S-nitrosylation

Nitrosylation of proteins is a reversible process similar to phosphorylation. This process is independent of cyclic guanosine monophosphate (cGMP) nitric oxide modification and a new way to regulate protein function, which is an important post-translational modification of proteins. Nitrosylation of proteins refers to the process by which nitric oxide (NO) groups are covalently attached to free sulfhydryl groups of specific cysteine residues in targeted proteins, thereby forming S-nitrosothiols (SNOs). S-nitrosylation modification widely exists in biological reactions, and can adjust biological processes by improving protein activity and protein interaction. The most widely used method for detecting protein S-nitrosylation is the biotinylation conversion method, the principle of which is to specifically reduce the modified SNO group to a sulfhydryl group and label it with biotin, which can be detected by western blotting way of detection.

S-nitrosation is a typical redox-dependent protein post-translational modification. This modification is a non-enzymatic reversible process and participates in many physiological processes (activation of defense genes, regulation of programmed cell death). S-nitrosation is an important protein post-translational modification method, which can regulate the mechanism, and it is of great significance for its efficient detection. Related services that Alfa Chemistry can provide include but not limited to:

• Efficient S-nitrosation detection
• Protein in vitro biotinylation
• Identify and quantify nitrosylation sites

Nitrosylation of proteins

NO forms nitrosothiols (-SNO) by reversibly covalently attaching to the free sulfhydryl groups of cysteine residues in proteins, a process known as nitrosylation of proteins. Modification by S-nitrosylation can improve protein subcellular localization, protein interaction, protein biological activity and protein stability. It is of great significance to efficiently detect S-nitrosylated target proteins and their interactions.

The advantages and features of the detection platform

• Specificity: Conversion of S-nitrosocysteine post-translational modification assays to use of antibodies
• Low signal-to-noise ratio: Professional biotinylated labels, low background
• Workflow compatibility: Compatibility of labeling methods with alternative analysis methods
• Easy and fast: Complete and easy to operate

Figure 2. Professional biotinylation platform, insisting on providing customers with high-quality services

Detection principle

• Block the free sulfhydryl groups on the sample protein
• The S-nitrososulfhydryl group on the sample protein is reduced to generate a free thiol covalently labeled with maleimide biotin
• Subsequent detection of biotin-labeled proteins with avidin-conjugated reagents

The future development

More and more studies have shown that the nitrosylation modification of proteins is involved in the regulation of the physiological processes of organisms, and the detection and identification of nitrosylation target proteins and target sites will greatly promote the development of research in this field . In the future, using new technologies and methods to identify more nitrosylated target proteins and target sites will be of great significance for revealing the process and mechanism of nitrosylation.

Alfa Chemistry takes advantage of competitive biotinylation services to provide biotin detection of protein S-nitrosylation and meet customers' needs with high-quality services. If you are interested in our services, please contact us immediately.

Reference:

1. Chen. L.C.; et al. Detection and analysis of plant protein S-nitrosylation modification. Chinese Bulletin of Botany. 2019, 54 (4): 497-502.

※ It should be noted that our service is only used for research, not for clinical use.