| 2014 |
LRRC8A (SWELL1) is an essential component of the volume-regulated anion channel (VRAC). Genome-wide RNAi screens independently identified LRRC8A as required for hypotonicity-induced iodide influx and VRAC currents. Genomic disruption of LRRC8A ablated VRAC currents, and point mutations in LRRC8A cause significant changes in VRAC anion selectivity, demonstrating that LRRC8A is a pore-forming component. LRRC8A forms heteromers with other LRRC8 family members (LRRC8B-E), and the isoform combination determines VRAC inactivation kinetics. Taurine flux and regulatory volume decrease also depend on LRRC8 proteins. |
Genome-wide siRNA screen, CRISPR genomic disruption, patch-clamp electrophysiology, taurine flux assay, point mutagenesis |
Science / Cell |
High |
24725410 24790029
|
| 2016 |
LRRC8 proteins together constitute the VRAC pore. SWELL1 and up to four other LRRC8 subunits assemble into heterogeneous complexes of ~800 kDa. When reconstituted into lipid bilayers, LRRC8 complexes are sufficient to form anion channels activated by osmolality gradients. Single-channel conductance depends on LRRC8 subunit composition. Low ionic strength in the absence of an osmotic gradient activates the complexes in bilayers, demonstrating that hypotonic stress can activate VRAC through a decrease in cytoplasmic ionic strength. |
Reconstitution into lipid bilayers, single-channel electrophysiology, size-exclusion chromatography (~800 kDa complex), patch-clamp |
Cell |
High |
26824658
|
| 2018 |
Cryo-EM and X-ray crystallography of homomeric LRRC8A reveal a hexameric channel architecture. The transmembrane domain is structurally related to connexin proteins, wide towards the cytoplasm but constricted on the extracellular side by a selectivity filter. An excess of basic residues in the filter and throughout the pore attracts anions by electrostatic interaction. The cytoplasmic leucine-rich repeat domain follows the transmembrane pore domain. |
Cryo-electron microscopy, X-ray crystallography |
Nature |
High |
29769723
|
| 2018 |
Cryo-EM structure of human LRRC8A shows a hexameric assembly. The transmembrane region features topology similar to gap junction channels. The LRR region (15 leucine-rich repeats) forms a long twisted arc. The channel pore is constricted on the extracellular side, where conserved polar and charged residues at the tip of the extracellular helix contribute to anion permeability. Two structural populations (compact and relaxed conformations) suggest that the LRR region undergoes rigid-body motions possibly implicated in pore opening. |
Single-particle cryo-electron microscopy |
Nature structural & molecular biology |
High |
30127360
|
| 2019 |
Cryo-EM structures of LRRC8A in lipid nanodiscs with the inhibitor DCPIB show that DCPIB plugs the channel in the extracellular selectivity filter, sterically occluding ion conduction. Constricted and expanded structures reveal coupled dilation of cytoplasmic LRRs and the channel pore, suggesting a gating mechanism by internal stimuli. Conformational differences between detergent and lipid bilayer structures demonstrate a critical role for the membrane lipid environment in determining channel structure, including intersubunit lipid binding sites. |
Single-particle cryo-electron microscopy, lipid nanodisc reconstitution, inhibitor-bound structure |
eLife |
High |
30775971
|
| 2018 |
The intracellular loop (IL) connecting TM2 and TM3 of LRRC8A and the first extracellular loop (EL1) of LRRC8C/D/E are both essential for VRAC activity. A 25-amino acid sequence unique to the LRRC8A IL is sufficient to generate homomeric VRAC activity when inserted into LRRC8C or LRRC8E. LRRC8 chimeras containing partial LRRC8A IL sequences exhibit altered anion permeability, rectification, and voltage sensitivity, indicating that the LRRC8A IL contributes to pore structure and function. |
Chimeric channel mutagenesis, patch-clamp electrophysiology in LRRC8-/- cells |
The Journal of general physiology |
High |
29853476
|
| 2018 |
The short N-terminal stretch preceding the first LRRC8 transmembrane domain determines VRAC conductance, ion permeability, and inactivation gating. Substituted-cysteine accessibility studies reveal that the first 15 LRRC8A residues are exposed to a hydrophilic environment. Glutamate 6 (E6) controls iodide-over-chloride permeability and voltage dependence of inactivation; restoring the negative charge by MTSES reverses these effects. Cd2+-mediated blocking data suggest the N termini come close together in the multimeric complex and may line the cytoplasmic pore. |
Substituted-cysteine accessibility method (SCAM), site-directed mutagenesis, patch-clamp, MTSES modification |
The Journal of biological chemistry |
High |
29925591
|
| 2023 |
2.8-Å cryo-EM structure of human LRRC8A shows well-resolved N-termini (NTs). The amino-terminal halves of NTs fold back into the pore and constrict the permeation path, forming a second selectivity filter working in series with the extracellular selectivity filter. The C-terminal halves of NTs interact with intracellular loops crucial for channel activation. Molecular dynamics simulations indicate that low ionic strength increases NT mobility and expands inter-helix distances, suggesting a mechanism for VRAC activation. |
Single-particle cryo-EM (2.8 Å), molecular dynamics simulations |
Cell reports |
High |
37543949
|
| 2022 |
Cryo-EM structures of heterohexameric LRRC8A:C channels reveal a predominant assembly with A:C ratio of 2:4 (four LRRC8A and two LRRC8C subunits). Four LRRC8A subunits cluster in their preferred closed-state conformation as pairs. The two LRRC8C subunits show greater flexibility, destabilizing the tightly packed A subunits and enhancing activation. Lipids embedded in the channel pore block ion conduction in the closed state. |
Single-particle cryo-EM, fiducial-tagging strategy for subunit identification, functional electrophysiology |
Nature structural & molecular biology / Nature structural & molecular biology |
High |
36522427 36928458
|
| 2014 |
LRRC8A is an indispensable component of the swelling-activated excitatory amino acid (EAA) release pathway in rat astrocytes. siRNA knockdown of LRRC8A dramatically reduced hypo-osmotic release of D-[3H]aspartate, L-glutamate, and taurine, and completely abolished ATP-stimulated release of EAAs and taurine from non-swollen astrocytes. |
siRNA knockdown, radiotracer efflux assays (D-[3H]aspartate, [14C]taurine), HPLC |
The Journal of physiology |
High |
25172945
|
| 2014 |
LRRC8A constitutively associates with the GRB2-GAB2 complex and lymphocyte-specific protein tyrosine kinase (LCK) in thymocytes. LRRC8A ligation activates AKT via the LCK-ZAP-70-GAB2-PI3K pathway, and AKT phosphorylation is markedly reduced in Lrrc8a-/- thymus. Thymic epithelial cells express an LRRC8A ligand critical for double-negative to double-positive thymocyte differentiation and survival in vitro. |
Co-immunoprecipitation, Lrrc8a-/- mouse phenotyping, bone marrow chimeras, flow cytometry, phospho-AKT immunoblot |
The Journal of experimental medicine |
Medium |
24752297
|
| 2017 |
SWELL1/LRRC8A regulates adipocyte insulin-PI3K-AKT2-GLUT4 signaling, glucose uptake, and lipid content via interactions of the SWELL1 C-terminal leucine-rich repeat domain (LRRD) with GRB2 and Cav1. Silencing GRB2 in SWELL1 KO adipocytes rescues insulin-pAKT2 signaling. In vivo, adipose-targeted SWELL1 KO reduces adiposity and impairs systemic glycemia and insulin sensitivity. |
Co-immunoprecipitation (LRRD-GRB2/Cav1 interaction), adipose-specific KO mice, patch-clamp, glucose uptake assay, siRNA rescue |
Nature cell biology |
High |
28436964
|
| 2020 |
LRRC8A/LRRC8E-containing VRACs transport cGAMP and cyclic dinucleotides across the plasma membrane, enabling cell-to-cell transmission of the innate immune second messenger cGAMP. Chemical blockade or genetic ablation of LRRC8A results in defective interferon responses to HSV-1. Enhancing VRAC activity by hypotonic swelling, cisplatin, GTPγS, or cytokines (TNF, IL-1) increases STING-dependent IFN responses to extracellular cGAMP. Lrrc8e-/- mice exhibit impaired IFN responses and compromised immunity to HSV-1. |
Biochemical transport assay, electrophysiology, genetic ablation (LRRC8A KO, Lrrc8e-/- mice), IFN ELISA, viral challenge |
Immunity |
High |
32277911
|
| 2020 |
LRRC8A forms complexes with LRRC8C and/or LRRC8E to transport cGAMP and other 2'3'-cyclic dinucleotides as an importer or exporter depending on the electrochemical gradient. LRRC8D inhibits cGAMP transport. Activation of LRRC8A channels by sphingosine 1-phosphate potentiates cGAMP transport. LRRC8A channels are the key cGAMP transporters in resting primary human vasculature cells. |
Genome-wide CRISPR screen, cGAMP transport assay, STING reporter assay, pharmacological activation/inhibition (S1P, DCPIB) |
Molecular cell |
High |
33171122
|
| 2019 |
Astrocytic SWELL1/LRRC8A mediates non-vesicular glutamate release through VRAC. Both cell swelling and receptor stimulation activate astrocytic VRAC, which requires SWELL1. Astrocyte-specific Swell1 KO mice exhibit impaired glutamatergic transmission (reduced presynaptic release probability and ambient glutamate), hippocampal-dependent learning/memory deficits, and attenuated neuronal excitability and brain damage after ischemic stroke. |
Astrocyte-specific conditional KO mice, whole-cell patch-clamp, glutamate release assay, behavioral testing, ischemia model (MCAO) |
Neuron |
High |
30982627
|
| 2018 |
SWELL1 mediates a swell-activated, depolarizing chloride current (ICl,SWELL) in murine and human β-cells. Hypotonic and glucose-stimulated β-cell swelling activates SWELL1-mediated ICl,SWELL, contributing to membrane depolarization and activation of voltage-gated Ca2+ channels (VGCC)-dependent intracellular calcium signaling. β-cell-targeted Swell1 KO mice have impaired glucose-stimulated insulin secretion and glucose tolerance. |
Patch-clamp electrophysiology, tamoxifen-inducible β-cell Swell1 KO mice, calcium imaging, insulin secretion assay, glucose tolerance test |
Nature communications |
High |
29371604 29773801
|
| 2021 |
Under hypertonic conditions, LRRC8A is phosphorylated and activated by MSK1 kinase downstream of the p38-MSK1 stress pathway. LRRC8A-mediated Cl- efflux then facilitates activation of the WNK kinase pathway, which promotes electrolyte influx via NKCC cotransporter for regulatory volume increase (RVI). The LRRC8A-S217A mutation impairs channel activation by MSK1, resulting in reduced RVI and cell survival. Identified by genome-wide CRISPR/Cas9 screen. |
Genome-wide CRISPR/Cas9 survival screen, site-directed mutagenesis (S217A), kinase assays, RVI volume measurements, NKCC inhibition |
Proceedings of the National Academy of Sciences |
High |
34083438
|
| 2016 |
LRRC8A physically co-localizes and co-immunoprecipitates with NADPH oxidase 1 (Nox1) and its p22phox subunit in vascular smooth muscle cells. LRRC8A is required for TNFα-induced extracellular superoxide production by Nox1, which in turn is essential for TNFR1 endocytosis, JNK phosphorylation, and NF-κB activation. LRRC8A siRNA reduces VRAC current and inhibits NF-κB-dependent inflammatory signaling. |
Co-immunoprecipitation, immunostaining co-localization, siRNA knockdown, superoxide assays (SOD inhibition), NF-κB reporter, receptor endocytosis assay |
Free radical biology & medicine |
Medium |
27838438
|
| 2021 |
LRRC8A physically interacts with NADPH oxidases Nox2, Nox4, and p22phox via its C-terminal leucine-rich repeat domain (LRRD). C-terminal LRRD mutant LRRC8A fails to co-immunoprecipitate with Nox2/Nox4/p22phox. LRRC8A knockdown suppresses AngII-induced ROS production, NADPH oxidase activity, and translocation of cytosolic subunits p47phox and p67phox, implicating LRRC8A-LRRD as the interface for Nox complex regulation in AngII-induced cardiac hypertrophy. |
Co-immunoprecipitation, domain-deletion mutant (LRRD), immunofluorescence co-localization, AAV9-siRNA in vivo knockdown, ROS/NADPH oxidase activity assays |
Free radical biology & medicine |
Medium |
33515753
|
| 2021 |
LRRC8A activates JAK2-STAT3 signaling via its C-terminal leucine-rich repeat domain (LRRD), which directly interacts with the adaptor protein GRB2. GRB2 is associated with and necessary for tyrosine-phosphorylated JAK2. This LRRC8A-GRB2-JAK2-STAT3 axis mediates TGF-β1-induced myofibroblast transformation and cardiac fibrosis following myocardial infarction. Myofibroblast-specific Lrrc8a KO attenuates fibrotic remodeling and ventricular dysfunction after MI. |
Co-immunoprecipitation, LRRD domain mutant, myofibroblast-specific conditional KO mice (periostin-Cre), RNA sequencing, immunoblot for JAK2/STAT3 phosphorylation |
Theranostics |
Medium |
35966575
|
| 2017 |
LRRC8A-containing VRAC in astrocytes shows subunit-dependent substrate specificity: LRRC8A/D-containing heteromers dominate release of uncharged osmolytes (taurine, myo-inositol), whereas LRRC8A/C/E-containing channels dominate release of charged osmolytes (D-aspartate). This demonstrates the existence of at least two distinct heteromeric VRACs in the same cell type. |
RNAi knockdown of individual LRRC8 subunits, radiotracer efflux assays (D-[14C]aspartate, [3H]taurine, myo-[3H]inositol) |
The Journal of physiology |
Medium |
28833202
|
| 2017 |
LRRC8A-LRRC8E heteromeric channels are dramatically activated (>10-fold) by oxidation of intracellular cysteine residues (by chloramine-T or tBHP), whereas LRRC8A-LRRC8C and LRRC8A-LRRC8D heteromers are strongly inhibited by oxidation. Endogenous VRAC currents in Jurkat T lymphocytes are inhibited by oxidation, consistent with their predominant LRRC8C/D expression. LRRC8 channel proteins are thus directly modulated by oxidation in a subunit-specific manner. |
Fluorescently-tagged constitutively active LRRC8 constructs, whole-cell patch-clamp, oxidant application (chloramine-T, tBHP), siRNA for subunit expression verification |
The Journal of physiology |
Medium |
28841766
|
| 2020 |
Intracellular LRRC8 proteins localize to lysosomes and generate lysosomal VRAC (Lyso-VRAC) currents in response to low cytoplasmic ionic strength. A double-leucine motif (L706L707) at the LRRC8A C-terminus is required for lysosomal targeting; mutation to alanines abolishes Lyso-VRAC but preserves plasma membrane VRAC. Lyso-VRAC is necessary for formation of lysosome-derived vacuoles that store and expel excess water. Selective elimination of Lyso-VRAC increases necrotic cell death under hypoosmotic, hypoxic, and hypothermic stress. |
Lysosome patch-clamp (whole-lysosome configuration), C-terminal targeting mutant (L706L707A), pharmacological tools, cell death assays |
Proceedings of the National Academy of Sciences |
High |
33139539
|
| 2018 |
Germ cell-specific disruption of Lrrc8a leads to abnormal sperm morphology (cytoplasm swelling, disorganized mitochondrial sheaths, angulated/coiled flagella) and male infertility in mice, consistent with impaired cell volume regulation in late spermatids. Sertoli cell-specific disruption does not cause infertility, indicating a cell-autonomous requirement in germ cells. |
Germ cell-specific and Sertoli cell-specific Lrrc8a KO mice, electron microscopy, sperm morphology analysis, fertility testing |
The Journal of biological chemistry |
High |
29880644 30135305
|
| 2022 |
NHE1 polarizes to the cell leading edge and SWELL1 polarizes to the cell trailing edge during confined migration. SWELL1 polarization confers migration direction and efficiency. Optogenetic RhoA activation at the cell front triggers SWELL1 redistribution and migration direction reversal in SWELL1-expressing but not SWELL1-knockdown cells. Dual NHE1/SWELL1 knockdown inhibits breast cancer extravasation and metastasis in vivo. |
Live-cell imaging, optogenetics (RhoA activation), siRNA knockdown, in vivo extravasation/metastasis model, mathematical modeling |
Nature communications |
Medium |
36253369
|
| 2021 |
Endothelial LRRC8A (SWELL1) is required for VRAC in HUVECs and regulates AKT-eNOS signaling under basal, stretch, and shear-flow conditions. LRRC8A forms a GRB2-Cav1-eNOS signaling complex. Endothelium-restricted Lrrc8a KO mice develop hypertension upon chronic angiotensin-II infusion and exhibit impaired retinal blood flow in type 2 diabetes. |
Co-immunoprecipitation (GRB2-Cav1-eNOS complex with LRRC8A), endothelium-specific KO mice, patch-clamp, flow/stretch experiments, blood pressure telemetry, retinal angiography |
eLife |
Medium |
33629656
|
| 2020 |
SWELL1 (LRRC8A) encodes a swell-activated anion channel in skeletal muscle cells that regulates PI3K-AKT, ERK1/2, and mTOR signaling, muscle differentiation, myoblast fusion, oxygen consumption, and glycolysis. LRRC8A overexpression in KO myotubes rescues myotube formation by boosting PI3K-AKT-mTOR signaling. Skeletal muscle-targeted KO mice have smaller myofibers, reduced force, decreased exercise endurance, increased adiposity, and glucose intolerance on high-fat diet. |
Skeletal muscle-specific Lrrc8a KO mice, LRRC8A overexpression rescue, patch-clamp, AKT/ERK/mTOR phosphoblot, myoblast fusion assay, metabolic/exercise phenotyping |
eLife |
Medium |
32930093
|
| 2019 |
LRRC8/VRAC channels are permeable to glutathione (GSH) with a PGSH/PCl ratio of ~0.1 under hypotonic conditions. LRRC8A KO cells show no GSH conductance. LRRC8/VRAC-mediated GSH efflux modulates intracellular ROS levels and contributes to TGFβ1-induced epithelial-to-mesenchymal transition (EMT); pharmacological (DCPIB) or siRNA inhibition of LRRC8A attenuates EMT by preserving intracellular GSH and reducing ROS. |
LRRC8A KO cells, GSH current measurement, intracellular GSH quantification, DCPIB inhibition, siRNA, EMT marker expression |
Cell death & disease |
Medium |
31804464
|
| 2016 |
LRRC8A downregulation in cisplatin-sensitive ovarian and alveolar carcinoma cells reduces p53 protein levels and downstream signaling (p21Waf1/Cip1, MDM2) and abolishes Caspase-9/-3 activation after cisplatin treatment. Cisplatin resistance correlates with reduced total LRRC8A expression (A2780) or reduced plasma membrane LRRC8A (A549). LRRC8A-dependent channel activity is upstream of cisplatin-induced apoptotic volume decrease. |
siRNA knockdown, pharmacological inhibition (NS3728, DIDS), immunoblot for p53/p21/MDM2/caspase activation, apoptosis assays |
American journal of physiology. Cell physiology |
Medium |
26984736
|
| 2016 |
Cisplatin accumulation in cells correlates with LRRC8A protein expression and VSOAC channel activity; cellular platinum content is high when VSOAC is activated and reduced when LRRC8A is silenced or pharmacologically inhibited. This demonstrates that LRRC8A-containing channels mediate cisplatin uptake. |
siRNA knockdown, ICP-MS platinum quantification, pharmacological inhibition, VRAC activation/inhibition |
Journal of inorganic biochemistry |
Medium |
27112899
|
| 2019 |
LRRC8/VRAC promotes mouse myoblast differentiation by facilitating plasma membrane hyperpolarization early during differentiation. LRRC8A knockdown or pharmacological VRAC inhibition reduces myogenin expression and suppresses myoblast fusion without affecting proliferation. VRAC acts upstream of K+ channel activation; inhibition prevents early hyperpolarization and the subsequent increase in intracellular steady-state Ca2+ levels during myogenesis. |
siRNA knockdown of LRRC8A, pharmacological VRAC inhibition, membrane potential measurement, Ca2+ imaging, myogenin immunostaining, myotube formation assay |
The Journal of biological chemistry |
Medium |
31387946
|
| 2022 |
In proximal tubules, LRRC8A and LRRC8D localize to basolateral membranes. Conditional deletion of LRRC8A in proximal (but not distal) tubules, and constitutive deletion of LRRC8D, cause proximal tubular injury, increased diuresis, and mild Fanconi-like symptoms. LRRC8C is exclusively found in vascular endothelium, and LRRC8E is specific for intercalated cells. These findings demonstrate that LRRC8A/D channels are required for basolateral exit of organic compounds in proximal tubules. |
Epitope-tagged LRRC8 knock-in mice (localization), conditional tubule-specific LRRC8A KO mice, constitutive LRRC8D KO, histology, urine/serum metabolomics |
Journal of the American Society of Nephrology |
High |
35777784
|
| 2020 |
LRRC8A is an essential regulator of hypotonicity-induced NLRP3 inflammasome activation in murine macrophages, but is dispensable for canonical DAMP-induced NLRP3 activation. Chemical, biochemical, and genetic (KO) approaches demonstrated this selective requirement. Canonical DAMP-dependent NLRP3 activation remains sensitive to chloride channel inhibitors, indicating additional chloride-sensing mechanisms. |
LRRC8A KO macrophages, VRAC pharmacological inhibitors, NLRP3 inflammasome activation assay (IL-1β/IL-18 secretion), ASC speck formation |
eLife |
Medium |
33216713
|
| 2024 |
Phosphorylation of LRRC8A at Serine 174 (S174) acts as a checkpoint for VRAC activity in the steady state. ATP-evoked K+ efflux via P2X receptors alleviates S174 phosphorylation, thereby activating VRAC and potentiating cGAMP transport. Mutagenesis of S174 modulates the ATP responsiveness of LRRC8A channels. |
Site-directed mutagenesis (S174), P2X receptor agonist/antagonist, K+ efflux assays, cGAMP transport assay, VRAC electrophysiology |
Journal of immunology |
Medium |
38847616
|
| 2023 |
LRRC8A channel inhibition in macrophages promotes phagocytosis by activating AMPK, inducing nuclear translocation of Nrf2, and increasing CD36 transcription. Conditional KO of Lrrc8a in macrophages accelerates hematoma clearance, reduces neuronal death, and improves functional recovery after intracerebral hemorrhagic stroke. |
Macrophage/microglia-specific Lrrc8a KO mice, AMPK inhibitor, Nrf2 nuclear translocation assay, CD36 expression, ICH mouse model, VRAC pharmacological inhibition |
iScience |
Medium |
36465125
|
| 2021 |
Brain-wide conditional deletion of LRRC8A causes fatal seizures in mice at 5–9 weeks of age. Hippocampal slice electrophysiology reveals increased pyramidal cell excitability and modified GABAergic inputs. LRRC8A-null hippocampi show decreased GLT-1, GAT-1, and glutamine synthetase protein levels, and reduced tissue glutamine, indicating that VRAC is required for normal astrocytic amino acid neurotransmitter homeostasis and brain excitability. |
NestinCre-driven brain-wide LRRC8A conditional KO, EEG/video seizure recording, brain slice patch-clamp, immunoblot, HPLC amino acid quantification |
FASEB journal |
Medium |
34469026
|
| 2003 |
A truncated form of LRRC8A (deletion of LRR7–9 at C-terminus), caused by chromosomal translocation, acts as a dominant negative to inhibit B cell development when expressed in murine bone marrow transplantation experiments. LRRC8A is expressed on T cells and B-lineage cells and is required for normal B cell development. |
Chromosomal translocation analysis, cDNA cloning, murine bone marrow transplantation with truncated LRRC8A expression, flow cytometry |
The Journal of clinical investigation |
Medium |
14660746
|
| 2018 |
GlialCAM/MLC1 modulates LRRC8/VRAC currents indirectly: MLC1 cannot potentiate VRAC when LRRC8A is knocked down, but LRRC8A and MLC1 do not co-localize or co-immunoprecipitate and MLC1 does not potentiate LRRC8-mediated VRAC currents in Xenopus oocytes. Lack of MLC1 increases phosphorylation of LRRC8C (a VRAC subunit), and MLC1 overexpression reduces ERK phosphorylation, suggesting indirect modulation through signal transduction pathways. |
Co-immunoprecipitation (negative), Xenopus oocyte expression, LRRC8A siRNA knockdown, ERK phosphorylation immunoblot, LRRC8C phosphorylation assay |
Neurobiology of disease |
Medium |
30076890
|
| 2023 |
LRRC8A associates with MPRIP (myosin phosphatase rho-interacting protein), confirmed by LRRC8A immunoprecipitation/mass spectrometry, confocal co-localization, proximity ligation assay, and IP/western blot. LRRC8A-MPRIP interaction links LRRC8A to RhoA-MYPT1-actin pathway; siLRRC8A or VRAC blockade decreases RhoA activity in VSMCs, and MYPT1 phosphorylation is reduced in VSMC-specific Lrrc8a KO mesenteries, contributing to enhanced vascular relaxation. |
Co-IP/mass spectrometry, proximity ligation assay, confocal imaging, VSMC-specific Lrrc8a KO mice, RhoA activity assay, MYPT1 phospho-immunoblot |
FASEB journal |
Medium |
37310356
|
| 2022 |
LRRC8A channel quantification in cells: approximately 10,000 VRAC channels per cell based on quantitative immunoblot with recombinant protein calibration. LRRC8A immunoprecipitation co-precipitates an excess of non-LRRC8A subunits, suggesting these subunits predominate numerically in heterohexamers. |
Quantitative immunoblot with recombinant protein calibration, co-immunoprecipitation of all five LRRC8 subunits from multiple tissues |
International journal of molecular sciences |
Low |
31771171
|
| 2024 |
BioID proximity labeling of LRRC8A identifies interactions with cell-cell junction proteins, calcium homeostasis regulators, kinases, and GTPase signaling components. Re-evaluation of LRRC8A in HCT116 LRRC8A-KO cells confirms no effect on cell proliferation or migration, consistent with PMID:31151189. |
BioID proximity-dependent biotinylation and mass spectrometry, LRRC8A-KO proliferation/migration assay |
Cell death discovery |
Low |
38909013
|