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Analysis of protein S-nitrosylation
Joan B Mannick1, Christopher M Schonhoff2
1University of Massachusetts Medical School, Worcester, Massachusetts.
Current Protocols in Protein Science
|April 23, 2008
Summary
S-nitrosylation, a reversible protein modification crucial for cell signaling, is challenging to study due to its easily disrupted bonds. Recent methods improve the detection of S-nitrosylated proteins in biological samples.
Area of Science:
- Biochemistry
- Cell Biology
- Chemical Biology
Background:
- S-nitrosylation is a post-translational modification involving the addition of a nitric oxide (NO) group to cysteine thiols.
- It functions as a critical on/off switch for protein activity in cellular signaling pathways.
- Unlike phosphorylation, S-nitrosylation occurs nonenzymatically, influenced by cellular redox conditions.
Purpose of the Study:
- To review the challenges associated with analyzing protein S-nitrosylation.
- To describe recently developed methods for measuring S-nitrosylated proteins in biological samples.
Main Methods:
- Discussion of the technical difficulties in detecting S-nitrosylation due to the labile S-NO bond.
- Overview of novel analytical techniques designed to overcome sample preparation and detection limits.
- Focus on methods suitable for biological samples.
Main Results:
- Protein S-nitrosylation analysis is hindered by the easily disrupted S-NO bond during sample preparation.
- The low abundance of S-nitrosylated proteins challenges current detection technologies.
- Several new methods have been developed to facilitate the measurement of protein S-nitrosylation.
Conclusions:
- Despite technical hurdles, advancements in analytical methods are improving the study of protein S-nitrosylation.
- These new techniques are essential for understanding the role of S-nitrosylation in cell signaling and redox biology.
- Further development in detection technologies is needed to fully elucidate the scope of S-nitrosylation.
Related Concept Videos
01:26Protein Modifications in the RER
Modification of secretory and transmembrane proteins entering the rough ER begins in the ER lumen. These modifications aid in protein folding and stabilize the acquired tertiary structure. Protein modifications in the rough ER co-occur at different stages of protein folding.
Broadly, these modifications can be categorized into four main categories — glycosylation, formation of disulfide bonds, assembly of protein subunits, and specific proteolytic cleavages like removal of signal sequences.
Broadly, these modifications can be categorized into four main categories — glycosylation, formation of disulfide bonds, assembly of protein subunits, and specific proteolytic cleavages like removal of signal sequences.
01:202° Amines to N-Nitrosamines: Reaction with NaNO2
Secondary amines react with nitrous acid to form N-nitrosamines, as depicted in Figure 1. Nitrous acid, a weak and unstable acid, is formed in situ from an aqueous solution of sodium nitrite and strong acids, such as hydrochloric acid or sulfuric acid, in cold conditions. In the presence of an acid, the nitrous acid gets protonated. The subsequent loss of water results in the formation of the electrophile known as nitrosonium ion.