Skywalker-TBC1D24 has a lipid-binding pocket mutated in epilepsy and required for synaptic function
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Mutations in TBC1D24 cause severe epilepsy and DOORS syndrome, but the molecular mechanisms underlying these pathologies are unresolved. We solved the crystal structure of the TBC domain of the Drosophila ortholog Skywalker, revealing an unanticipated cationic pocket conserved among TBC1D24 homologs. Cocrystallization and biochemistry showed that this pocket binds phosphoinositides phosphorylated at the 4 and 5 positions. The most prevalent patient mutations affect the phosphoinositide-binding pocket and inhibit lipid binding. Using in vivo photobleaching of Skywalker-GFP mutants, including pathogenic mutants, we showed that membrane binding via this pocket restricts Skywalker diffusion in presynaptic terminals. Additionally, the pathogenic mutations cause severe neurological defects in flies, including impaired synaptic-vesicle trafficking and seizures, and these defects are reversed by genetically increasing synaptic PI(4,5)P 2 concentrations through synaptojanin mutations. Hence, we discovered that a TBC domain affected by clinical mutations directly binds phosphoinositides through a cationic pocket and that phosphoinositide binding is critical for presynaptic function.
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The TBC1D7 apo structure reveals an architecture highly conserved with other TBC domain proteins, with little conformational difference in comparison to the TSC1-complexed structures.
“The apo structure (Fig. 5D) reveals an architecture highly conserved with other TBC domain proteins (Fischer et al., 2016; Fukuda, 2011; Park et al., 2011), with little conformational difference in comparison to the TSC1-complexed structures.”
From Signal integration in the (m)TORC1 growth pathway (2018) · cites this paper as Fischer et al., 2016
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- Authors
- Fischer, B., Lüthy, K., Paesmans, J., De Koninck, C., Maes, I., Swerts, J., Kuenen, S., Uytterhoeven, V., Verstreken, P., & Versées, W.
- Affiliations
- Vrije Universiteit Brussel (BE)
- VIB-KU Leuven Center for Brain & Disease Research (BE)
- Corresponding author
- Verstreken, Patrik
- Versées, Wim
- ORCID
- Venue
- Nature Structural & Molecular Biology
- Published
- 2016-09-26
- Volume
- 23
- Issue
- 11
- Pages
- 965-973
- ISSN
- 1545-9985 (ISSN-L)
- 1545-9993
- OpenAlex
- W2524717647
- Cited by
- 62
- References
- 63
- Citations by year
- 2026: 2 · 2025: 2 · 2024: 8 · 2023: 3 · 2022: 6 · 2021: 5 · 2020: 13 · 2019: 5 · 2018: 10 · 2017: 6 · 2016: 1 · 2015: 1
- FWCI
- 5.05
- Type
- journal article
- Language
- en
- Open access
- green
- zenodo.org
- Licence
- other-oa
- www.springer.com/tdm (tdm)
- Locations
- Nature Structural & Molecular Biology (publishedVersion)
- PubMed (publishedVersion)
- VUBIR (Vrije Universiteit Brussel) (submittedVersion)
- Zenodo (CERN European Organization for Nuclear Research) (submittedVersion, other-oa)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Health Sciences › Medicine › Psychiatry and Mental health › Epilepsy research and treatment
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetics and Neurodevelopmental Disorders
- Keywords
- TBC1D24, early-onset epilepsy, TBC domain, phosphoinositide binding, lipid-binding pocket, phosphatidylinositol 4,5-bisphosphate, synaptic vesicle trafficking, presynaptic terminals, protein diffusion, synaptojanin, photobleaching, Drosophila
- MeSH terms
- Protein Domains, Animals, Binding Sites, Carrier Proteins, Diffusion, Drosophila melanogaster, Epilepsy, Hand Deformities, Congenital, Hearing Loss, Sensorineural, Humans, Membrane Proteins, Intellectual Disability, Models, Molecular, Mutation, Nails, Malformed, Nerve Tissue Proteins, Phosphatidylinositols, Phosphoric Monoester Hydrolases, Protein Conformation, Synaptic Vesicles, Crystallography, X-Ray, Craniofacial Abnormalities, GTPase-Activating Proteins, rab GTP-Binding Proteins, Drosophila Proteins
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