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

Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

7.9K
Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
7.9K
Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

13.8K
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...
13.8K
Phosphorylation01:02

Phosphorylation

52.2K
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...
52.2K
Protein Modifications in the RER01:26

Protein Modifications in the RER

5.8K
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...
5.8K
Disassembly of Intermediate Filaments01:35

Disassembly of Intermediate Filaments

2.2K
Intermediate filaments (IFs) do not undergo spontaneous disassembly. Enzymes, kinases, and phosphatases add and remove phosphates from specific sites to regulate their disassembly. The IF concentration in the cytoplasm also regulates the disassembly. If the concentration crosses a threshold, it activates the protein kinases in the vicinity, allowing the phosphorylation of IFs.
Keratin proteins, found at the cell periphery near cell junctions, undergo a cycle of assembly and disassembly. In Type...
2.2K
Notch Signaling Pathway03:14

Notch Signaling Pathway

4.6K
The Notch signaling pathway is a major intracellular signaling pathway that is highly conserved over a broad spectrum of metazoan species. It stands unique from other intracellular signaling mechanisms in animals because notch protein itself acts as the receptor as well as the primary signaling molecule.
The Notch gene came into the limelight in 1914 after the discovery that its mutation in Drosophila melanogaster leads to a serrated (or "notched") wing margin phenotype. It was not...
4.6K

You might also read

Related Articles

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

Sort by
Same author

Functional comparison of SP6 RNA polymerase and T7 RNA polymerase.

PloS one·2026
Same author

Human RNA ligase 1 as a novel regulator of ribosome function and translation under oxidative stress.

Nucleic acids research·2026
Same author

Immobilization of Endonuclease A from Serratia marcescens on cellulose membranes.

Biotechnology letters·2026
Same author

Incidence of Naloxone-Use Before and After Implementing Opioid Safety Measures for Patients Hospitalised on General Wards in a Swiss University Hospital.

Drug safety·2026
Same author

Quantifying Ligand-to-Protein Distances in Complex Environments Using Intermolecular <sup>19</sup>F PRE NMR Spectroscopy.

Chembiochem : a European journal of chemical biology·2026
Same author

Recommendations for perioperative pain medicine focusing on structures, processes, and organisation.

Swiss medical weekly·2026

Related Experiment Video

Updated: Oct 17, 2025

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
12:26

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

Published on: May 3, 2018

18.9K

Structural and functional consequences of NEDD8 phosphorylation.

Katrin Stuber1,2,3, Tobias Schneider2,3, Jill Werner1,2

  • 1Dept. of Biology, University of Konstanz, Konstanz, Germany.

Nature Communications
|October 13, 2021
PubMed
Summary
This summary is machine-generated.

Phosphorylation of ubiquitin (Ub) and NEDD8 at S65 impacts their structure and function, activating Parkin. Phosphorylated NEDD8 interacts with HSP70, highlighting the importance of these modifications.

More Related Videos

Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins
12:47

Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins

Published on: December 27, 2016

19.0K
Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

8.6K

Related Experiment Videos

Last Updated: Oct 17, 2025

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
12:26

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

Published on: May 3, 2018

18.9K
Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins
12:47

Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins

Published on: December 27, 2016

19.0K
Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

8.6K

Area of Science:

  • Molecular and Cellular Biology
  • Biochemistry
  • Post-Translational Modifications

Background:

  • Ubiquitin (Ub) and Ub-like proteins (Ubls) like NEDD8 are key post-translational modifiers.
  • While phosphorylated Ub (pUb) functions are known, Ubl phosphorylation effects remain unclear.
  • Phosphorylation is a critical regulatory mechanism for protein function.

Purpose of the Study:

  • To investigate the consequences of NEDD8 phosphorylation at S65.
  • To compare the effects of NEDD8 phosphorylation with those of Ub phosphorylation.
  • To explore the functional implications of phosphorylated NEDD8 (pNEDD8) interactions.

Main Methods:

  • Analysis of NEDD8 phosphorylation at S65 and its effect on structural dynamics.
  • Assessment of allosteric activation of the Ub ligase Parkin by pUb and pNEDD8.
  • Characterization of protein interactomes for unmodified and phosphorylated Ub/NEDD8.
  • Investigation of pNEDD8 interaction with HSP70 family members and its effect on ATPase activity.

Main Results:

  • NEDD8 phosphorylation at S65 mirrors Ub phosphorylation, affecting structural dynamics similarly.
  • Both pUb and pNEDD8 allosterically activate the Parkin Ub ligase.
  • pNEDD8 exhibits distinct interactomes compared to unmodified NEDD8, notably increased interaction with HSP70 family members.
  • pNEDD8 more significantly stimulates HSP70 ATPase activity than unmodified NEDD8.

Conclusions:

  • Phosphorylation of Ub and NEDD8 at S65 is functionally significant.
  • The functions of pUb and pNEDD8 do not necessitate covalent attachment to target proteins.
  • Phosphorylation of Ubls represents a crucial layer of regulatory control.