Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Nitric Oxide Signaling Pathway01:28

Nitric Oxide Signaling Pathway

6.5K
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...
6.5K
Phosphorylation01:02

Phosphorylation

55.2K
The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
55.2K
Phosphorylation01:02

Phosphorylation

7.8K
7.8K
Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

15.4K
Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
15.4K
Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

4.6K
4.6K
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

19.0K
When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze...
19.0K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Corrigendum to: "NO modulates human airway smooth muscle function by altering glucose-6-phosphate dehydrogenase effects on sGC function in asthma" [Redox Biology 95 (2026) 104262].

Redox biology·2026
Same author

NO modulates human airway smooth muscle function by altering glucose-6-phosphate dehydrogenase effects on sGC function in asthma.

Redox biology·2026
Same author

NO modulates human airway smooth muscle function by altering glucose-6-phosphate dehydrogenase effects on sGC function in asthma.

bioRxiv : the preprint server for biology·2026
Same author

Myoglobin leaching into the serum of IDA mice is associated with elevated sGC activation under anemic conditions that correlates with increased myoglobin expression.

Nitric oxide : biology and chemistry·2026
Same author

Arginine substitution of conserved Lys<sup>609</sup> and Lys<sup>733</sup> impairs FMN dynamics, electron transfer, and nitric oxide production in endothelial nitric oxide synthase.

International journal of biological macromolecules·2026
Same author

Distinct catalytic activity of Staphylococcus Aureus nitric oxide synthase compared to other bacterial NOS-like enzymes.

Journal of inorganic biochemistry·2026

Related Experiment Video

Updated: Mar 15, 2026

Application of Genetically Encoded Fluorescent Nitric Oxide (NO&#8226;) Probes, the geNOps, for Real-time Imaging of NO&#8226; Signals in Single Cells
08:32

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

Published on: March 16, 2017

13.4K

Phosphorylation Controls Endothelial Nitric-oxide Synthase by Regulating Its Conformational Dynamics.

Mohammad Mahfuzul Haque1, Sougata Sinha Ray2, Dennis J Stuehr3

  • 1From the Department of Pathobiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195 haquem@ccf.org.

The Journal of Biological Chemistry
|September 11, 2016
PubMed
Summary

Phosphorylation of endothelial NO synthase (eNOS) at Ser1179 enhances its activity by altering electron transfer and reductase domain dynamics. This finding clarifies how Ser1179 phosphorylation regulates eNOS function.

Keywords:
catalysiseNOSelectron transferfast reaction kineticsflavoproteinheme reductionkinetic modelnitric oxide synthasepre-steady-state kineticssimulation

More Related Videos

Platelet-based Detection of Nitric Oxide in Blood by Measuring VASP Phosphorylation
07:13

Platelet-based Detection of Nitric Oxide in Blood by Measuring VASP Phosphorylation

Published on: January 7, 2019

8.5K
Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

9.0K

Related Experiment Videos

Last Updated: Mar 15, 2026

Application of Genetically Encoded Fluorescent Nitric Oxide (NO&#8226;) Probes, the geNOps, for Real-time Imaging of NO&#8226; Signals in Single Cells
08:32

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

Published on: March 16, 2017

13.4K
Platelet-based Detection of Nitric Oxide in Blood by Measuring VASP Phosphorylation
07:13

Platelet-based Detection of Nitric Oxide in Blood by Measuring VASP Phosphorylation

Published on: January 7, 2019

8.5K
Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

9.0K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Enzyme Kinetics

Background:

  • Endothelial NO synthase (eNOS) activity is regulated by calmodulin (CaM) binding and phosphorylation.
  • Phosphorylation at Ser1179 is a key regulatory event that enhances eNOS activity in response to physiological stimuli.
  • The precise mechanism by which Ser1179 phosphorylation increases eNOS activity remains unclear.

Purpose of the Study:

  • To investigate the mechanistic impact of Ser1179 phosphorylation on eNOS activity.
  • To determine how the phosphomimetic mutation S1179D affects electron flux and reductase domain conformation.
  • To compare the effects of S1179D mutation with CaM binding on eNOS activity.

Main Methods:

  • Stopped-flow spectroscopy to monitor reaction kinetics.
  • Computer modeling to analyze conformational dynamics.
  • Site-directed mutagenesis to create the S1179D phosphomimetic mutant.

Main Results:

  • The S1179D mutation in CaM-free eNOS accelerated flavin reduction and altered reductase domain conformational equilibrium and transitions.
  • These effects were comparable in magnitude to those induced by CaM binding to wild-type eNOS.
  • Combined S1179D mutation and CaM binding resulted in more pronounced changes in reductase domain parameters.

Conclusions:

  • Ser1179 phosphorylation, mimicked by the S1179D mutation, directly enhances eNOS activity by modulating electron flux and reductase domain dynamics.
  • These changes within the reductase domain are sufficient to explain the observed stimulation of both reductase and NO synthase activities.
  • This study elucidates a key regulatory mechanism of eNOS and provides a basis for studying other phosphorylation events.