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Related Concept Videos

2° Amines to N-Nitrosamines: Reaction with NaNO201:20

2° 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.
Nitrosation of Enols01:19

Nitrosation of Enols

The nitrosation reaction is one of the methods of preparing 1,2-diketones. The enol tautomer of the starting ketone reacts with sodium nitrite in hydrochloric acid, generating the 1,2-diketone after hydrolysis.
Rate-Determining Steps03:08

Rate-Determining Steps

Relating Reaction Mechanisms
In a multistep reaction mechanism, one of the elementary steps progresses significantly slower than the others. This slowest step is called the rate-limiting step (or rate-determining step). A reaction cannot proceed faster than its slowest step, and hence, the rate-determining step limits the overall reaction rate.
The concept of rate-determining step can be understood from the analogy of a 4-lane freeway with a short-stretch of traffic-bottleneck caused due to...
Electrophilic Aromatic Substitution: Nitration of Benzene01:20

Electrophilic Aromatic Substitution: Nitration of Benzene

The nitration of benzene is an example of an electrophilic aromatic substitution reaction. It involves the formation of a very powerful electrophile, the nitronium ion, which is linear in shape. The reaction occurs through the interaction of two strong acids, sulfuric and nitric acid.
Inorganic Nitrogen Assimilation01:22

Inorganic Nitrogen Assimilation

Nitrogen is an essential element in biological systems, forming a crucial component of proteins, nucleic acids, and other cellular constituents. Many bacteria and archaea acquire nitrogen in the form of nitrate (NO₃⁻) or ammonia (NH₃), which are then assimilated into biomolecules through specific enzymatic pathways.Assimilatory Nitrate ReductionWhen nitrate enters the cell, it undergoes a two-step reduction process known as assimilatory nitrate reduction. Initially, the enzyme nitrate reductase...
Preparation of Amines: Reduction of Oximes and Nitro Compounds01:29

Preparation of Amines: Reduction of Oximes and Nitro Compounds

Oximes can be reduced to primary amines using catalytic hydrogenation, hydride reduction, or sodium metal reduction. The reduction of aliphatic and aromatic nitro compounds to primary amines takes place by either catalytic hydrogenation or by using active metals like Fe, Zn, and Sn in the presence of an acid.
Though catalytic hydrogenation can reduce nitrobenzenes, the reduction is nonselective in the presence of other functional groups. For instance, if nitrobenzene contains an aldehyde group,...

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Assessment of the nitrosylation process.

Didier Borderie1, Ohvanesse G Ekindjian

  • 1Laboratoire de biochimie A, Hôpital Cochin APHP, 27 rue du faubourg Saint Jacques, France. didier.borderie@cch.aphp.fr

Current Opinion in Clinical Nutrition and Metabolic Care
|August 8, 2008
PubMed
Summary

Accurately measuring protein S-nitrosylation is crucial for understanding its role in cell signaling. Current popular methods like chemiluminescence and biotin switch assays have limitations and require careful controls for reliable results.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Physiology

Background:

  • Protein S-nitrosylation is a critical post-translational modification.
  • Nitric oxide (NO) signaling pathways are vital in cellular regulation.
  • Understanding S-nitrosylation is key to deciphering NO-mediated cellular functions.

Purpose of the Study:

  • To review and analyze methodologies for measuring S-nitrosylated proteins.
  • To identify the principles and limitations of current protein S-nitrosylation assays.
  • To provide insights into the accurate quantification of S-nitrosylated proteins in biological samples.

Main Methods:

  • Review of chemiluminescence-based assays.
  • Analysis of biotin switch assay protocols.
  • Evaluation of sample pretreatment effects on S-nitrosylation.
  • Discussion of detection limits and assay sensitivity.

Main Results:

  • Chemiluminescence and biotin switch assays are widely adopted but possess potential pitfalls.
  • Many assays operate near their detection limits, impacting accuracy.
  • Sample preparation can inadvertently alter or cleave the S-NO bond.
  • Quantitative validation using orthogonal methods is often necessary.

Conclusions:

  • Protein S-nitrosylation is an important NO-regulated signaling mechanism.
  • Existing measurement techniques require critical assessment and validation.
  • Implementing additional controls and employing multiple methodologies ensures reliable S-nitrosylation data.