| 2003 |
GlyR-gephyrin binding is dependent on the presence of an intact C-terminal MoeA homology (E-domain) of gephyrin. The N10Y missense mutation and alternative splicing of GPHN transcripts do not disrupt GlyR-gephyrin interactions or collybistin-induced cell-surface clustering. |
Yeast two-hybrid, functional clustering assay, RT-PCR isoform analysis, mutagenesis |
The Journal of biological chemistry |
Medium |
12684523
|
| 2003 |
Gephyrin (encoded by GEPH/GPHN) is required during molybdenum cofactor assembly for insertion of molybdenum into the cofactor; loss-of-function mutations abrogate all molybdoenzyme activities. |
Human mutation analysis combined with biochemical phenotyping of molybdoenzyme activities in patients |
Human mutation |
Medium |
12754701 21031595
|
| 2001 |
Gephyrin functions as a peripheral membrane scaffolding protein anchoring glycine receptors to subsynaptic microtubules; it also plays a role in GABA-A receptor localization at the synapse and in molybdenum cofactor biosynthesis, as established by knockout mouse phenotypes. |
Knockout mouse analysis, gene structure determination, localization studies |
Gene |
Medium |
11418245
|
| 2001 |
In a translocation t(11;14)(q23;q24)-associated leukemia, the GPHN C-terminal half (including a tubulin-binding site and MoeA homology domain) is fused to MLL AT-hook and DNA methyltransferase homology domains, generating an MLL-GPHN fusion protein. Genomic breakpoint analysis identified topoisomerase-II DNA-binding sites spanning both breakpoints, suggesting VP16/topoisomerase-II-induced double-strand breaks and non-homologous end joining as the generation mechanism. |
cDNA library screening, fusion transcript identification, genomic breakpoint analysis |
Genes, chromosomes & cancer |
Medium |
11579461
|
| 2024 |
Gephyrin assembles with heteromeric α2β glycine receptors (GlyRs) into micron-sized clusters at the plasma membrane. Neuroligin-2 further increases cluster sizes and GlyR concentration. A positively charged N-terminus sequence of the GlyR β subunit is essential for glycine affinity modulation through clustering. Ligand re-binding to adjacent clustered GlyRs alters kinetics but not chemical equilibrium. |
Heterologous expression clustering assay, electrophysiology, mutagenesis of GlyR β subunit N-terminus |
bioRxivpreprint |
Medium |
bio_10.1101_2024.10.17.618879
|
| 2024 |
In zebrafish, Gephyrin (Gphnb) is enriched in myelin on GABAergic and glycinergic axons. Loss of gphnb causes longer myelin sheaths specifically on GABAergic axons and shifts myelin placement toward glutamatergic axons at the expense of GABAergic axons, indicating gephyrin mediates selective axon-class-dependent myelination by oligodendrocytes. |
Zebrafish gphnb loss-of-function genetics, imaging of myelin sheath length and distribution per axon class |
bioRxivpreprint |
Medium |
bio_10.1101_2024.10.02.616365
|
| 2025 |
Cryo-EM combined with biochemical reconstitution and mutational analyses shows that full-length gephyrin forms a stable dimer as the basic oligomeric unit, which further assembles into linear and oblique tetramers and linear hexamers. A critical segment of the flexible central linker adopts two distinct conformations, one of which occludes the receptor-binding site, and this segment harbors key phosphorylation sites, providing a mechanistic link between phosphorylation state, linker conformation, and receptor binding. |
Cryo-electron microscopy, biochemical reconstitution, mutational analysis |
bioRxivpreprint |
High |
bio_10.1101_2025.09.01.673457
|
| 2025 |
Collybistin induces gephyrin self-oligomerization into a high-molecular-weight (>5 MDa) gephyrin-collybistin complex at GABAergic synapses. Plasma-membrane phosphoinositides promote complex formation and are critical for membrane targeting and stabilization. Gephyrin phosphorylation at Ser325 abolishes complex formation with collybistin, impairing collybistin-dependent gephyrin clustering at GABAergic synapses. |
Biochemical reconstitution of gephyrin-collybistin complex, phosphorylation site mutagenesis (Ser325), phosphoinositide binding assays, synaptic clustering assays |
bioRxivpreprint |
High |
bio_10.1101_2025.01.20.633899
|
| 2025 |
WNK1 kinase (and its effector SPAK) directly phosphorylates two previously uncharacterized residues in the central linker region of gephyrin. This phosphorylation controls GABA-A receptor synaptic diffusion, clustering, membrane stability, and endocytosis at inhibitory synapses. Activation of WNK signaling stabilizes GABA-A Rs at inhibitory synapses; inhibition enhances receptor internalization. Expression of a phospho-mimetic form of gephyrin at WNK-targeted sites produces anxiolytic effects in vivo. |
Phosphorylation site identification and mutagenesis, live imaging of receptor diffusion, electrophysiology of GABAergic currents, in vivo behavioral assay (anxiety) |
bioRxivpreprint |
Medium |
bio_10.1101_2025.09.05.674425
|
| 2025 |
A knock-in mouse model of the gephyrin microdeletion Δ199-233 (within the C-domain, removing a region that includes the S-palmitoylation site at Cys212) disrupts synaptic targeting of gephyrin in dissociated hippocampal neurons. Despite unexpectedly facilitating receptor interaction in vitro, inhibitory signal transmission is reduced. Compensatory changes occur at excitatory synapses (smaller but more numerous PSD-95 clusters). Loss of the C-domain palmitoylation site is required for correct synaptic targeting. |
Knock-in mouse model, immunofluorescence of synaptic clustering in hippocampal neurons, electrophysiology of inhibitory transmission, PSD-95 cluster analysis |
bioRxivpreprint |
Medium |
bio_10.1101_2025.08.26.672322
|
| 2024 |
Gephyrin, the main inhibitory receptor scaffold, is organized into sub-synaptic domains (SSDs) in vivo with distinct nanoscale arrangements depending on subcellular location and presynaptic partner. Chronic chemogenetic increases in cortical activity cause a reduction in gephyrin SSD volume specifically at axo-axonic (but not axo-dendritic) synapses, functionally weakening those contacts, demonstrating activity-dependent nanoscale remodeling of the inhibitory scaffold. |
dSTORM super-resolution microscopy in vivo, chemogenetic manipulation, electrophysiological measurement of synaptic strength |
bioRxivpreprint |
Medium |
bio_10.1101_2024.11.29.625981
|
| 2024 |
An E3 ligase-dependent tool (paGFE3) that targets the RING domain of Mdm2 to gephyrin degrades gephyrin protein and ablates inhibitory synapses in response to 400 nm light, confirming that gephyrin is required for maintenance of inhibitory synaptic structure. |
Photoactivatable optogenetic degradation system (paGFE3), immunofluorescence of gephyrin levels and inhibitory synapse markers |
bioRxivpreprint |
Low |
bio_10.1101_2024.09.23.614589
|