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- EMDB-60798: Cryo-EM structure of human TRPV4 intracellular domain in complex ... -
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Open data
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Basic information
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Title | Cryo-EM structure of human TRPV4 intracellular domain in complex with GTPase RhoA | |||||||||
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![]() | Complex / MEMBRANE PROTEIN / Hydrolase | |||||||||
Function / homology | ![]() stretch-activated, monoatomic cation-selective, calcium channel activity / blood vessel endothelial cell delamination / regulation of response to osmotic stress / osmosensor activity / vasopressin secretion / positive regulation of striated muscle contraction / calcium ion import into cytosol / negative regulation of brown fat cell differentiation / positive regulation of microtubule depolymerization / positive regulation of macrophage inflammatory protein 1 alpha production ...stretch-activated, monoatomic cation-selective, calcium channel activity / blood vessel endothelial cell delamination / regulation of response to osmotic stress / osmosensor activity / vasopressin secretion / positive regulation of striated muscle contraction / calcium ion import into cytosol / negative regulation of brown fat cell differentiation / positive regulation of microtubule depolymerization / positive regulation of macrophage inflammatory protein 1 alpha production / hyperosmotic salinity response / positive regulation of chemokine (C-X-C motif) ligand 1 production / positive regulation of chemokine (C-C motif) ligand 5 production / cartilage development involved in endochondral bone morphogenesis / alpha-beta T cell lineage commitment / aortic valve formation / mitotic cleavage furrow formation / positive regulation of lipase activity / bone trabecula morphogenesis / endothelial tube lumen extension / skeletal muscle satellite cell migration / positive regulation of vascular associated smooth muscle contraction / angiotensin-mediated vasoconstriction involved in regulation of systemic arterial blood pressure / SLIT2:ROBO1 increases RHOA activity / RHO GTPases Activate Rhotekin and Rhophilins / Roundabout signaling pathway / negative regulation of intracellular steroid hormone receptor signaling pathway / Axonal growth inhibition (RHOA activation) / Axonal growth stimulation / cellular hypotonic salinity response / cleavage furrow formation / regulation of neural precursor cell proliferation / cellular hypotonic response / cortical microtubule organization / regulation of modification of postsynaptic actin cytoskeleton / regulation of osteoblast proliferation / forebrain radial glial cell differentiation / multicellular organismal-level water homeostasis / regulation of modification of postsynaptic structure / apical junction assembly / cell junction assembly / negative regulation of cell migration involved in sprouting angiogenesis / cellular response to chemokine / establishment of epithelial cell apical/basal polarity / positive regulation of vascular permeability / beta selection / regulation of systemic arterial blood pressure by endothelin / osmosensory signaling pathway / negative regulation of oxidative phosphorylation / negative regulation of motor neuron apoptotic process / RHO GTPases Activate ROCKs / negative regulation of cell size / RHO GTPases activate CIT / cell-cell junction assembly / Sema4D induced cell migration and growth-cone collapse / PCP/CE pathway / cellular response to osmotic stress / positive regulation of monocyte chemotactic protein-1 production / RHO GTPases activate KTN1 / calcium ion import / cell volume homeostasis / positive regulation of podosome assembly / apolipoprotein A-I-mediated signaling pathway / positive regulation of alpha-beta T cell differentiation / Sema4D mediated inhibition of cell attachment and migration / wound healing, spreading of cells / positive regulation of leukocyte adhesion to vascular endothelial cell / PI3K/AKT activation / Wnt signaling pathway, planar cell polarity pathway / motor neuron apoptotic process / odontogenesis / ossification involved in bone maturation / regulation of aerobic respiration / regulation of focal adhesion assembly / TRP channels / negative chemotaxis / apical junction complex / cortical actin cytoskeleton / EPHA-mediated growth cone collapse / myosin binding / stress fiber assembly / positive regulation of cytokinesis / regulation of neuron projection development / diet induced thermogenesis / positive regulation of macrophage chemotaxis / RHOC GTPase cycle / cellular response to cytokine stimulus / cerebral cortex cell migration / ERBB2 Regulates Cell Motility / microtubule polymerization / cleavage furrow / semaphorin-plexin signaling pathway / calcium ion import across plasma membrane / androgen receptor signaling pathway / ficolin-1-rich granule membrane / RHOA GTPase cycle / mitotic spindle assembly / negative regulation of cell-substrate adhesion / alpha-tubulin binding / Rho protein signal transduction Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.16 Å | |||||||||
![]() | Yuan Z / Ruan SS / Li SL | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Inactivation of RhoA for Hypertension Treatment Through the TRPV4-RhoA-RhoGDI1 Axis. Authors: Jiawen Wang / Zhen Yuan / Na Yu / Qian Jiao / Honglei Zhou / Wenjie Liao / Jiwei Shan / Shanshan Ruan / Yi Zhao / Ya Mo / Luyao Qi / Tiejun Li / Jianjun Fu / Bowen Ke / Yufang Xu / Xuhong ...Authors: Jiawen Wang / Zhen Yuan / Na Yu / Qian Jiao / Honglei Zhou / Wenjie Liao / Jiwei Shan / Shanshan Ruan / Yi Zhao / Ya Mo / Luyao Qi / Tiejun Li / Jianjun Fu / Bowen Ke / Yufang Xu / Xuhong Qian / Jian Zhang / Zhenjiang Zhao / Shiliang Li / Rui Wang / Honglin Li Abstract: BACKGROUND: The RhoA (Ras homolog family member A) signaling pathway is pivotal in regulating vascular smooth muscle cells (VSMCs) function and blood pressure homeostasis. Current inhibitors of the ...BACKGROUND: The RhoA (Ras homolog family member A) signaling pathway is pivotal in regulating vascular smooth muscle cells (VSMCs) function and blood pressure homeostasis. Current inhibitors of the RhoA signaling pathway are limited in hypertension treatment, suffering from poor efficacy, insufficient specificity, and developmental challenges. METHODS: Cryo-electron microscopy (EM), proximity ligation assay (PLA), and site-directed mutagenesis were used to explore the mechanism of RhoA activity regulation. VSMC, hypertensive animal models, ...METHODS: Cryo-electron microscopy (EM), proximity ligation assay (PLA), and site-directed mutagenesis were used to explore the mechanism of RhoA activity regulation. VSMC, hypertensive animal models, and Myh11-CRE (smooth muscle-specific RhoGDI1 knockout) mice were used to investigate the role of the TRPV4 (transient receptor potential cation channel subfamily V member 4)-RhoA-RhoGDI1 (Rho GDP dissociation inhibitor 1) axis in hypertension. RESULTS: AH001 ((R)-1-(3-ethylphenyl) ethane-1,2-diol) was identified as a novel inhibitor of the RhoA signaling pathway. It targets the TRPV4-RhoA-RhoGDI1 axis to effectively sequester inactive RhoA- ...RESULTS: AH001 ((R)-1-(3-ethylphenyl) ethane-1,2-diol) was identified as a novel inhibitor of the RhoA signaling pathway. It targets the TRPV4-RhoA-RhoGDI1 axis to effectively sequester inactive RhoA-GDP in the plasma membrane and cytoplasm, which is distinct from typical RhoA inhibition modes. The cryo-EM structure of the TRPV4-RhoA complex showed that AH001-bound TRPV4 adopts a closed state with RhoA in an inactive GDP-bound state. Functional studies further revealed that AH001 reduced the pool of active RhoA by enhancing TRPV4-RhoA binding and facilitating RhoGDI1-RhoA interaction in VSMC. This inhibition notably decreased both acute and long-term blood pressure and prevented vascular remodeling in Ang II-induced hypertensive mice and spontaneously hypertensive rats. However, these antihypertensive effects were weakened in and Myh11-CRE mice. Additionally, AH001 effectively inhibited VSMC contraction via the RhoA/ROCK (Rho-associated protein kinase)/MYPT1 (myosin phosphatase target subunit 1)/MLC (myosin light chain 2) signaling pathway and suppressed VSMC phenotype switching to myofibroblasts through the RhoA/ROCK/LIMK1 (LIM domain kinase)/cofilin/MRTF-A (myocardin-related transcription factor A)/SRF (serum response factor) signaling cascade. TRPV4 and RhoGDI1 knockdown attenuated AH001's inhibition of VSMC contraction and phenotypic switching to myofibroblasts. CONCLUSIONS: This study revealed a novel mode of RhoA signaling inhibition targeting the TRPV4-RhoA-RhoGDI1 axis, offering new insights for future antihypertensive drug development and proposing ...CONCLUSIONS: This study revealed a novel mode of RhoA signaling inhibition targeting the TRPV4-RhoA-RhoGDI1 axis, offering new insights for future antihypertensive drug development and proposing innovative strategies for targeting challenging Rho GTPases. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 118 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 19 KB 19 KB | Display Display | ![]() |
Images | ![]() | 58.4 KB | ||
Filedesc metadata | ![]() | 6.4 KB | ||
Others | ![]() ![]() | 115.9 MB 115.9 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 911 KB | Display | ![]() |
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Full document | ![]() | 910.7 KB | Display | |
Data in XML | ![]() | 13.9 KB | Display | |
Data in CIF | ![]() | 16.5 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 9iqyMC ![]() 9iqxC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.08 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: #1
File | emd_60798_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_60798_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Cryo-EM structure of human TRPV4 intracellular domain in complex ...
Entire | Name: Cryo-EM structure of human TRPV4 intracellular domain in complex with GTPase RhoA |
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Components |
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-Supramolecule #1: Cryo-EM structure of human TRPV4 intracellular domain in complex ...
Supramolecule | Name: Cryo-EM structure of human TRPV4 intracellular domain in complex with GTPase RhoA type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: Transient receptor potential cation channel subfamily V member 4
Macromolecule | Name: Transient receptor potential cation channel subfamily V member 4 type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 73.038719 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: VFNRPILFDI VSRGSTADLD GLLPFLLTHK KRLTDEEFRE PSTGKTCLPK ALLNLSNGRN DTIPVLLDIA ERTGNMREFI NSPFRDIYY RGQTALHIAI ERRCKHYVEL LVAQGADVHA QARGRFFQPK DEGGYFYFGE LPLSLAACTN QPHIVNYLTE N PHKKADMR ...String: VFNRPILFDI VSRGSTADLD GLLPFLLTHK KRLTDEEFRE PSTGKTCLPK ALLNLSNGRN DTIPVLLDIA ERTGNMREFI NSPFRDIYY RGQTALHIAI ERRCKHYVEL LVAQGADVHA QARGRFFQPK DEGGYFYFGE LPLSLAACTN QPHIVNYLTE N PHKKADMR RQDSRGNTVL HALVAIADNT RENTKFVTKM YDLLLLKCAR LFPDSNLEAV LNNDGLSPLM MAAKTGKIGI FQ HIIRREV TDEDTRHLSR KFKDWAYGPV YSSLYDLSSL DTCGEEASVL EILVYNSKIE NRHEMLAVEP INELLRDKWR KFG AVSFYI NVVSYLCAMV IFTLTAYYQP LEGTPPYPYR TTVDYLRLAG EVITLFTGVL FFFTNIKDLF MKKCPGVNSL FIDG SFQLL YFIYSVLVIV SAALYLAGIE AYLAVMVFAL VLGWMNALYF TRGLKLTGTY SIMIQKILFK DLFRFLLVYL LFMIG YASA LVSLLNPCAN MKVCNEDQTN CTVPTYPSCR DSETFSTFLL DLFKLTIGMG DLEMLSSTKY PVVFIILLVT YIILTF VLL LNMLIALMGE TVGQVSKESK HIWKLQWATT ILDIERSFPV FLRKAFRSGE MVTVGKSSDG TPDRRWCFRV DEVNWSH UniProtKB: Transient receptor potential cation channel subfamily V member 4 |
-Macromolecule #2: Transforming protein RhoA
Macromolecule | Name: Transforming protein RhoA / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO / EC number: small monomeric GTPase |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 21.799158 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MAAIRKKLVI VGDGACGKTC LLIVFSKDQF PEVYVPTVFE NYVADIEVDG KQVELALWDT AGQEDYDRLR PLSYPDTDVI LMCFSIDSP DSLENIPEKW TPEVKHFCPN VPIILVGNKK DLRNDEHTRR ELAKMKQEPV KPEEGRDMAN RIGAFGYMEC S AKTKDGVR ...String: MAAIRKKLVI VGDGACGKTC LLIVFSKDQF PEVYVPTVFE NYVADIEVDG KQVELALWDT AGQEDYDRLR PLSYPDTDVI LMCFSIDSP DSLENIPEKW TPEVKHFCPN VPIILVGNKK DLRNDEHTRR ELAKMKQEPV KPEEGRDMAN RIGAFGYMEC S AKTKDGVR EVFEMATRAA LQARRGKKKS GCLVL UniProtKB: Transforming protein RhoA |
-Macromolecule #3: GUANOSINE-5'-DIPHOSPHATE
Macromolecule | Name: GUANOSINE-5'-DIPHOSPHATE / type: ligand / ID: 3 / Number of copies: 1 / Formula: GDP |
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Molecular weight | Theoretical: 443.201 Da |
Chemical component information | ![]() ChemComp-GDP: |
-Macromolecule #4: MAGNESIUM ION
Macromolecule | Name: MAGNESIUM ION / type: ligand / ID: 4 / Number of copies: 1 / Formula: MG |
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Molecular weight | Theoretical: 24.305 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOCONTINUUM (6k x 4k) / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 64000 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.0 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |