Related Experiment Video

Updated: Jun 9, 2026

Comparative Strategies for Ubiquitination Detection in Mammalian Cell Lysates Using SMAD2/SMURF2 as a Model
09:00

Comparative Strategies for Ubiquitination Detection in Mammalian Cell Lysates Using SMAD2/SMURF2 as a Model

Published on: April 17, 2026

The WW-HECT protein Smurf2 interacts with the Docking Protein NEDD9/HEF1 for Aurora A activation

Finola E Moore1, Evan C Osmundson1, Jennifer Koblinski2,3

  • 1Department of Molecular Pharmacology and Biological Chemistry, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.

Cell Division
|September 10, 2010
PubMed

Insights

The HECT-WW protein Smurf2 stabilizes NEDD9 (also known as HEF1/Cas-L), a key protein in cell cycle progression and cancer metastasis. Smurf2 depletion delays mitotic entry by inhibiting Aurora A kinase activation.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Cancer Research

Background:

  • The adaptor protein NEDD9 (HEF1/Cas-L) is crucial for regulating cell motility, invasion, cell cycle, and cancer metastasis.
  • NEDD9 localizes to the centrosome and is essential for Aurora A kinase activation during mitosis.

Purpose of the Study:

  • To investigate the interaction between Smurf2 and NEDD9.
  • To elucidate the role of Smurf2 in NEDD9 protein stability and its impact on mitotic entry.

Main Methods:

  • Co-immunoprecipitation assays to confirm physical association between Smurf2 and NEDD9.
  • Western blotting to assess protein levels of NEDD9 following Smurf2 manipulation.
  • Analysis of mitotic entry and Aurora A kinase activation in Smurf2-depleted cells.

Main Results:

  • Smurf2 physically associates with NEDD9, promoting its polyubiquitination and proteasomal degradation, thus regulating NEDD9 stability.
  • Smurf2 depletion significantly reduces NEDD9 protein levels and impairs Aurora A kinase activation at the G2/M phase.
  • Overexpression of Smurf2 leads to increased endogenous NEDD9 protein levels.

Conclusions:

  • The Smurf2-NEDD9 complex is vital for maintaining NEDD9 protein stability.
  • This interaction is critical for the timely activation of Aurora A kinase, ensuring proper entry into mitosis.

More Related Videos

Real-Time Monitoring of Aurora kinase A Activation using Conformational FRET Biosensors in Live Cells06:29

Real-Time Monitoring of Aurora kinase A Activation using Conformational FRET Biosensors in Live Cells

Published on: July 30, 2020

Identifying Protein-protein Interaction in Drosophila Adult Heads by Tandem Affinity Purification (TAP)10:36

Identifying Protein-protein Interaction in Drosophila Adult Heads by Tandem Affinity Purification (TAP)

Published on: December 5, 2013

Related Concept Videos

Tail-anchoring of Proteins in the ER Membrane01:45

Tail-anchoring of Proteins in the ER Membrane

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...
Export of Misfolded Proteins out of the ER01:32

Export of Misfolded Proteins out of the ER

After folding, the ER assesses the quality of secretory and membrane proteins. The correctly folded proteins are cleared by the calnexin cycle for transport to their final destination, while misfolded proteins are held back in the ER lumen. The ER chaperones attempt to unfold and refold the misfolded proteins but sometimes fail to achieve the correct native conformation. Such terminally misfolded proteins are then exported to the cytosol by ER-associated degradation or ERAD pathway for...
Hedgehog Signaling Pathway02:33

Hedgehog Signaling Pathway

The Hedgehog gene (Hh) was first discovered due to its control of the growth of disorganized, hair-like bristles phenotype in Drosophila, much like hedgehog spines. Hh plays a crucial role in the development of organs and the maintenance of homeostasis in both invertebrates and vertebrates. However, while Drosophila has only one Hh protein, mammals have multiple functional Hedgehog proteins - Sonic (Shh), Desert (Dhh), and Indian Hedgehog (Ihh). All of these homologous proteins have adapted to...
Anaphase Promoting Complex00:50

Anaphase Promoting Complex

The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...
Assembly of Signaling Complexes01:30

Assembly of Signaling Complexes

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,...
Coat Assembly and GTPases01:33

Coat Assembly and GTPases

Vesicles incorporate different coat protein subunits in different cell locations, which changes the properties of the coat, such as the shape and geometry of the transport vesicles. Thus, vesicle coat proteins also play a significant role in cargo selection.
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...