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Structural determinants for high-affinity binding in a Nedd4 WW3* domain-Comm PY motif complex.
Voula Kanelis1, M Christine Bruce, Nikolai R Skrynnikov
1Programme in Structural Biology and Biochemistry, Hospital for Sick Children, Toronto, Ontario M5G 1X8, Canada.
Structure (London, England : 1993)
|March 15, 2006
Summary
The WW3* domain of Drosophila Nedd4 interacts extensively with Commissureless, revealing new insights into neural development and protein regulation.
Area of Science:
- Molecular Biology
- Neuroscience
- Structural Biology
Background:
- The WW domains of Drosophila Nedd4 (dNedd4) and Commissureless (Comm) PY motifs are crucial for axon guidance and muscle development.
- Understanding the precise molecular interactions is key to elucidating developmental processes.
Purpose of the Study:
- To determine the solution structure of the dNedd4 WW3* domain in complex with a Commissureless PY motif.
- To investigate the molecular basis of WW domain-ligand recognition and its role in dNedd4-mediated ubiquitination.
Main Methods:
- Solution structure determination using NMR spectroscopy.
- Site-directed mutagenesis to probe protein-protein interactions.
- Biochemical binding assays to quantify interaction affinities.
Main Results:
- The WW3* domain interacts with residues both N- and C-terminal to the canonical PY motif of Comm.
- A helical turn formed by Comm residues Y232'-L236' engages with WW3*, expanding known WW domain-ligand interaction interfaces.
- A variable loop in WW3* is identified as critical for high-affinity binding.
Conclusions:
- The extensive interaction network between dNedd4 WW3* and Comm provides novel insights into WW domain-ligand recognition.
- These findings clarify the regulation of Comm ubiquitination and internalization, essential for central nervous system and muscle development.