2026-10-14T19:27:22Z
https://api.figshare.com/v2/oai
oai:figshare.com:article/34049536
2026-10-01T17:47:05Z
category_4
category_12
category_13
category_14
category_15
category_16
category_64
category_128
portal_5
item_type_1
month_year_10_2026
<p>EGL-2(G574E) depends on the potassium selectivity filter and the intrinsic ligand.</p>
Xinyu Huang (679541)
Sudharsan Kannan (25158064)
Gail A. Robertson (15007385)
Han Wang (254423)
Biochemistry
Cell Biology
Genetics
Molecular Biology
Neuroscience
Physiology
Cancer
Hematology
regulate neuronal excitability
promoting neuron ala
influence channel gating
hallmark cyclic nucleotide
genetic analysis demonstrated
forward genetics reveals
electrophysiological analysis showed
dependent channel activation
calcium imaging showed
binding homology domains
adjacent channel subunits
accelerates activation kinetics
subfamily kcnh channels
conserved gating mechanism
important sleep regulator
findings identify egl
div >< p
caenorhabditis elegans </
similarly suppresses sleep
2 functions cell
elegans </
suppresses sleep
kcnh voltage
vivo </
conserved cnbhd
central sleep
shifts voltage
second substitution
negative potentials
intrinsic ligand
g574r ),
function effects
corresponding residue
<p>(<b>A</b>) Conservation of the potassium selectivity filter in EAG channels. Top panel: amino acid sequence alignment of the selectivity filter region of rEAG1, dEAG, and EGL-2. The red arrowhead marks the conserved phenylalanine (F) residue; its substitution with alanine (F456A) in dEAG abolishes potassium currents. Bottom panel: side views of the selectivity filter formed by two adjacent subunits of rEAG1 (PDB:5K7L) or EGL-2 (predicted by AlphaFold3). (<b>B</b>) Structural model of the PAS-CNBHD interaction in EGL-2, predicted by AlphaFold3. The PAS domain and CNBHD shown are from adjacent subunits. The intrinsic ligand motif (Y686N687L688), located near the end of CNBHD, is important for PAS-CNBHD interaction. (<b>C-D</b>) Quantification of sleep behavior shown as the fraction of quiescent worms for the indicated genotypes and conditions. All strains contain the transgene (HS::EGF) for conditional overexpression of EGF via 15-min heat shock (HS) at 33 °C to induce sleep. Pre and post refer to before and 2 h after heat shock, respectively. (C) EGL-2(F452A), the equivalent of the dEAG(F456A) mutation that abolishes potassium currents, was introduced into the endogenous <i>egl-2(tan200[G574E])</i> locus. (D) The intrinsic ligand motif (Y686N687L688) of EGL-2 was mutated to ANA in the endogenous <i>egl-2(tan200[G574E])</i> locus. Each dot represents one independent assay containing 15–25 animals on a single plate. Mean ± SEM are shown. ****, p < 0.0001. One-way ANOVA with Dunnett’s multiple-comparisons test for post-heat shock samples, comparing each genotype to <i>egl-2(G574E)</i> mutants.</p>
2026-10-01T17:54:06Z
Image
Figure
10.1371/journal.pgen.1012255.g005
https://figshare.com/articles/figure/_p_EGL-2_G574E_depends_on_the_potassium_selectivity_filter_and_the_intrinsic_ligand_p_/34049536
CC BY 4.0