S-Nitrosylation Analysis

S-nitrosylation (SNO) refers to the covalent attachment of a nitroso group (NO) to the thiol group (S) of specific cysteine residues within proteins, forming S-nitrosoproteins. Thiol groups belong to a subset of cysteine residues in proteins, and the resulting SNOs are S-nitrosoproteins. The half-life of SNOs in the cytoplasm is very short because there are many reductases, including glutathione (GSH) and thioredoxins, that denitrosylate proteins. Therefore, SNOs are usually stored in the plasma membrane, vesicles, stroma, and folding of lipophilic proteins to protect them from denitrosylation. For example, apoptosis-mediating caspases are stored in the form of SNOs in the mitochondrial intermembrane space.

The process of S-nitrosylation is reversible. Denitrosylation is an enzyme-catalyzed process that reverses the S-nitrosylation process. However, S-nitrosylation is not a random event and only specific cysteine residues undergo S-nitrosylation. Abnormal S-nitrosylation may lead to protein misfolding, synaptic damage, and cell apoptosis. Dysfunction of SNO signaling is also associated with the pathogenesis of many diseases, such as Alzheimer's and cardiovascular diseases.

S-nitrosylation

Due to the presence of multiple cysteines in proteins and the unstable nature of SNOs, it is difficult to detect S-nitrosylated cysteines and distinguish them from non-S-nitrosylated amino acids. Biotech company Biopike has established a highly sensitive analytical platform capable of analyzing S-nitrosylated cysteines in both eukaryotes and prokaryotes. Additionally, we have optimized detection processes to provide faster and more sensitive S-nitrosylated cysteine analysis services.

General workflow of S-nitrosylation analysis services

1. Hydrolysis of proteins in gel or solution;
2. Enrichment of S-nitrosylated cysteine using anti-S-nitrosylated cysteine antibodies;
3. HPLC separation followed by ESI-TOF MS/MS analysis;
4. Mass spectrometry data analysis.

Technical platform

Ion chromatography

High-performance liquid chromatography (HPLC)

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS)

Related services

Post-translational modification proteomics analysis
Histone post-translational modification analysis
Protein SUMOylation identification
Protein disulfide bond identification and quantification