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

Protein Translocation Machinery on the ER Membrane01:28

Protein Translocation Machinery on the ER Membrane

7.2K
The translocon complex situated on the ER membrane is the main gateway for the protein secretory pathway. It facilitates the transport of nascent peptides into the ER lumen and their insertion into the ER membrane.
Sec61 protein conducting channel
In eukaryotes, the translocon complex comprises a core heterotrimeric translocator channel called the Sec61 complex. This channel includes three transmembrane proteins, Sec61α, Sec61β, and Sec61γ, and is the largest subunit of the...
7.2K
Assembly of Signaling Complexes01:30

Assembly of Signaling Complexes

6.9K
Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
6.9K
Enzyme-linked Receptors01:00

Enzyme-linked Receptors

88.0K
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...
88.0K
Tail-anchoring of Proteins in the ER Membrane01:45

Tail-anchoring of Proteins in the ER Membrane

4.0K
Tail-anchored, or TA, proteins are estimated to make up to 3-5% of membrane proteins found in the eukaryotic cell. Such proteins have a single transmembrane domain located approximately 30 amino acid residues upstream from the C-terminal end. As a result, the signal recognition particle (SRP) cannot guide a TA protein to the ER membrane for cotranslational insertion. Hence, they are integrated into the ER membrane post-translationally using their C-terminal end as the anchor. TA proteins...
4.0K

You might also read

Related Articles

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

Sort by
Same author

Kinesin-1 conformational dynamics are controlled by a cargo-sensitive TPR switch.

eLife·2026
Same author

The E3-ome gene-centric compendium reveals the human E3 ligase landscape.

Cell·2026
Same author

Structural basis of isethionate transport by a TRAP transporter from a sulfate-reducing bacterium.

Structure (London, England : 1993)·2025
Same author

Unconventional structure and function of PHD domains from additional sex combs-like proteins.

The FEBS journal·2025
Same author

DET1 dynamics underlie cooperative ubiquitination by CRL4<sup>DET1-COP1</sup> complexes.

Science advances·2025
Same author

On the function of TRAP substrate-binding proteins: the isethionate-specific binding protein IseP.

The Biochemical journal·2024

Related Experiment Video

Updated: Mar 10, 2026

Assaying Protein Kinase Activity with Radiolabeled ATP
08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

19.5K

Structural Studies of ERK2 Protein Complexes.

Johannes F Weijman1, Stefan J Riedl2, Peter D Mace3

  • 1Biochemistry Department, Otago School of Medical Sciences, University of Otago, 56, 710 Cumberland St., Dunedin, 9054, New Zealand.

Methods in Molecular Biology (Clifton, N.J.)
|December 8, 2016
PubMed
Summary

Researchers developed methods to crystallize ERK2-PEA-15 complexes. This work aids in understanding how regulators bind to the ERK1/2 signaling pathway

Keywords:
Activation loopCrystallizationD-PeptideDEF-dockingDEJLERK1ERK2MAPKPEA-15

More Related Videos

Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy
09:16

Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy

Published on: September 11, 2015

8.8K
Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting
10:08

Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting

Published on: December 9, 2022

2.7K

Related Experiment Videos

Last Updated: Mar 10, 2026

Assaying Protein Kinase Activity with Radiolabeled ATP
08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

19.5K
Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy
09:16

Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy

Published on: September 11, 2015

8.8K
Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting
10:08

Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting

Published on: December 9, 2022

2.7K

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Cell Signaling

Background:

  • Extracellular signal-regulated kinases (ERK1/2) are key kinases in the RAS-RAF-MEK-ERK pathway.
  • ERK1/2 interact with substrates and regulators via D-peptide and DEF-docking sites.
  • Understanding of DEF-docking site interactions is less developed compared to D-peptide interactions.

Purpose of the Study:

  • To describe methods for obtaining crystallization-grade complexes of ERK2-PEA-15.
  • To facilitate structural studies of ERK1/2 regulators interacting with the activation loop.
  • To advance understanding of PEA-15's role in regulating ERK kinase activity.

Main Methods:

  • Protein complex purification and characterization.
  • Crystallization techniques for protein-protein complexes.
  • Structural biology approaches to analyze kinase-regulator interactions.

Main Results:

  • Successful derivation of crystallization-grade ERK2-PEA-15 complexes.
  • PEA-15 engages both D-peptide and DEF-docking sites on ERK2.
  • PEA-15 sequesters the ERK2 activation loop across different phosphorylation states.

Conclusions:

  • The described methods are effective for producing stable ERK2-PEA-15 complexes for structural analysis.
  • These methods can be adapted for studying other ERK1/2 regulators.
  • This work provides a foundation for detailed structural insights into ERK1/2 regulation.