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Related Concept Videos

Nitric Oxide Signaling Pathway01:28

Nitric Oxide Signaling Pathway

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Nitric oxide (NO), an inorganic gas, acts as a potent second messenger in most animal and plant tissues. NO diffuses out of the cells that produce it and enters the neighboring cells to generate a downstream response. NO synthase (NOS) catalyzes NO production by the deamination of the amino acid arginine. There are three isoforms of NOS. Endothelial cells have endothelial NOS (eNOS), nerve and muscle cells have neuronal NOS (nNOS), and macrophages produce inducible NOS (iNOS) upon exposure...
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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.
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2° Amines to N-Nitrosamines: Reaction with NaNO201:20

2° Amines to N-Nitrosamines: Reaction with NaNO2

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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.
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1° Amines to Diazonium or Aryldiazonium Salts: Diazotization with NaNO2 Mechanism

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Nitrous acid is a relatively weak and unstable acid prepared in situ by the reaction of sodium nitrite and cold, dilute hydrochloric acid. In an acidic solution, the nitrous acid undergoes protonation when it loses water to form a nitrosonium ion—an electrophile. Nitrous acid reacts with primary amines to give diazonium salts. The reaction is called diazotization of primary amines.
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Enzyme-linked Receptors01:00

Enzyme-linked Receptors

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Enzyme-linked receptors are proteins that act as both receptor and enzyme, activating multiple intracellular signals. This is a large group of receptors that include the receptor tyrosine kinase (RTK) family. Many growth factors and hormones bind to and activate the RTKs.
Neurotrophin (NT) receptors are a family of RTKs, including trkA, trkB, and trkC (tropomyosin-related kinase) receptors. TrkA is specific for nerve growth factor (NGF), neurotrophin-6, and neurotrophin-7. TrkB binds...
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Preparation of Amines: Reduction of Oximes and Nitro Compounds01:29

Preparation of Amines: Reduction of Oximes and Nitro Compounds

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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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Application of Genetically Encoded Fluorescent Nitric Oxide (NO&#8226;) Probes, the geNOps, for Real-time Imaging of NO&#8226; Signals in Single Cells
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Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells

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Engineering nitric oxide synthase chimeras to function as NO dioxygenases.

Zhi-Qiang Wang1, Mohammad Mahfuzul Haque2, Katherine Binder2

  • 1Department of Chemistry and Biochemistry, Kent State University Geauga, Burton, OH 44021, United States.

Journal of Inorganic Biochemistry
|March 26, 2016
PubMed
Summary
This summary is machine-generated.

Researchers engineered nitric oxide synthase (NOS) chimeras to study NO dioxygenase activity. One chimera, V346I, showed significantly increased NO dioxygenase activity, consuming more NO than it released.

Keywords:
CatalysisChimeraElectron transferHeme reductionKoxStopped-flow

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Analytical Techniques for Assaying Nitric Oxide Bioactivity
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Area of Science:

  • Biochemistry
  • Enzymology
  • Molecular Biology

Background:

  • Nitric oxide synthases (NOSs) are enzymes catalyzing l-arginine oxidation to nitric oxide (NO) and l-citrulline.
  • NOS isoforms (iNOS, nNOS, eNOS) share structural similarity but differ in NO production kinetics.
  • A competing NO dioxygenase reaction can consume NO, impacting its physiological signaling.

Purpose of the Study:

  • To investigate the catalytic mechanisms of NOS enzymes, particularly their NO dioxygenase activity.
  • To design and characterize novel NOS chimeras with altered NO dioxygenase versus NO release functions.
  • To elucidate the structural and kinetic basis for differential NO dioxygenase activity in NOS.

Main Methods:

  • Construction of three NOS chimeras: iNOSoxy/nNOSred (WT), V346I iNOSoxy/nNOSred, and iNOSoxy/S1412D nNOSred.
  • Enzymatic assays to measure NO release and NO dioxygenase activity of the chimeras.
  • Kinetic parameter determination and computational modeling of NOS catalytic behavior.

Main Results:

  • The V346I chimera exhibited significantly reduced NO release compared to wild-type (WT) and S1412D chimeras.
  • The V346I chimera demonstrated a markedly higher NO dioxygenase activity, consuming 2-4 molecules of NO per molecule released.
  • WT and S1412D chimeras showed nearly equivalent NO release and NO dioxygenase activities.
  • Computer simulations accurately mimicked experimental outcomes, providing insights into catalytic mechanisms.

Conclusions:

  • Engineered NOS chimeras can be used to dissect the balance between NO synthesis and degradation.
  • Specific mutations, like V346I, can dramatically shift NOS function towards NO dioxygenation.
  • Understanding these catalytic behaviors is crucial for modulating NO bioavailability in biological systems.