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Proteomics to Identify Proteins Interacting with P2X2 Ligand-Gated Cation Channels
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Role of Ribeye PXDLS/T-binding cleft in normal synaptic ribbon function
Jie Zhu1, Caixia Lv1, Diane Henry2
1Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06520.
Biorxiv : the Preprint Server for Biology
|January 3, 2024
Summary
The PXDLS/T-binding cleft of Ribeye is crucial for recruiting it to synaptic ribbons and ensuring normal synaptic function in auditory and vestibular systems. Mutations disrupt this binding, impairing synaptic transmission and ribbon structure.
Area of Science:
- Neuroscience
- Cell Biology
- Auditory and Vestibular Systems
Background:
- Sensory hair cells in auditory and vestibular systems use ribbon synapses for high-fidelity signal encoding.
- Ribeye, a key synaptic ribbon protein, has a B-domain similar to CtBP2, featuring a binding cleft for PXDLS/T motifs.
Approach:
- Investigated the function of Ribeye's PXDLS/T-binding cleft using cell culture (INS-cells, HEK cells) and zebrafish models.
- Utilized overexpressed wild-type and mutant Ribeye-YFP, and Piccolo fragments to assess protein interactions and synaptic function.
Key Points:
- The PXDLS/T-binding cleft of Ribeye interacts with proteins like Piccolo, which possess the PVDLT motif.
- Mutations in this binding cleft prevent Ribeye's proper localization to synaptic ribbons.
- Restoring wild-type Ribeye in zebrafish rescued synaptic transmission and morphology, while mutant Ribeye caused abnormal aggregates and failed to rescue function.
Conclusions:
- The PXDLS/T-binding cleft is essential for Ribeye recruitment to synaptic ribbons.
- This interaction is critical for maintaining normal synaptic structure and function in sensory hair cells.
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