TRPV4 expresses in bone cell lineages and TRPV4-R616Q mutant causing Brachyolmia in human reveals “loss-of-interaction” with cholesterol

Das, Rashmita; Goswami, Chandan

doi:10.1016/j.bbrc.2019.07.042

Cited by 23 publications

(17 citation statements)

References 19 publications

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“…Allosteric sites in vanilloid TRP isoforms may include Cholesterol Recognition/Interaction Amino acid Consensus (CRAC)-like (KDLFRFLL) recognition motifs that span Loop 4 – TM5 (28, 82, 84). Consistent with our results, loss of TRPV4-cholesterol interaction in the TRPV4 R616Q CARC mutation was associated with gain-of-function for TRPV4 (67). It remains to be seen whether cholesterol influences the context of the cellular sensory response.…”

Section: Discussionsupporting

confidence: 92%

“…Interestingly, Cav-1 precipitated a small portion of the ∼75kDa variant, suggesting that TRPV4 variants might be differentially susceptible to caveolar interactions. Non-raft low- cholesterol regions reduce membrane mobility of TRPV4, with loss of cholesterol interaction resulting in gain-of-function for the channel (67). Similar results were observed for TRPM8, which shows enhanced gating in non-raft domains (50).…”

Section: Discussionmentioning

confidence: 99%

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Membrane cholesterol regulates TRPV4 function, cytoskeletal expression, and the cellular response to tension

Lakk

Hoffmann

Gorusupudi

et al. 2020

Preprint

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Cholesterol plays important roles in the regulation of membrane viscoelasticity, protein function and cellular health but despite its association with debilitating pressure-related diseases its role in mechanotransduction is not well understood. Here, we investigated how alterations in membrane cholesterol levels impact activation of TRPV4 (transient receptor potential vanilloid isoform 4), a stretch-activated cation channel, in trabecular meshwork (TM) cells that control intraocular pressure in the mammalian eye. Human TM cells responded to pharmacological depletion of free membrane cholesterol with augmented TRPV4 activation by agonist GSK1016790A, swelling and strain, and by increased membrane expression of TRPV4 immunoreactivity. Cyclic stretch increased the membrane cholesterol/phosphatidylcholine ratio and upregulated expression of F-actin, with the low-cholesterol effects reversed by cholesterol supplementation. Cholesterol depletion induced a constitutive current in TRPV4-expressing Xenopus laevis oocytes and obviated its activation by hypotonic swelling. Taken together, these findings suggest that cholesterol regulates translocation and activation of TRPV4, and guides its participation in macromolecular complexes, cytoskeletal architecture and cell-matrix contacts; in turn membrane cholesterol levels are dynamically regulated by the biomechanical milieu. By regulating TM pressure sensitivity, calcium homeostasis and contractility, cholesterol-TRPV4 interactions could influence IOP homeostasis and its role in glaucoma and diabetes.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Membrane cholesterol regulates TRPV4 function, cytoskeletal expression, and the cellular response to tension

Lakk

Hoffmann

Gorusupudi

et al. 2020

Preprint

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“…Allosteric sites may include cholesterol recognition/interaction amino acid consensus-like (KDLFRFLL) recognition motifs that span loop 4-TM5 of TRPV4 ( 30 , 88 , 89 ). Consistent with our results, loss of TRPV4-cholesterol interaction in the TRPV4 R616Q inverted cholesterol recognition/interaction amino acid consensus motif mutation was associated with gain of function for TRPV4 ( 90 ). Cholesterol thus appears to regulate the context of the cellular sensory response such that TRPV4 activity in yeast (which cannot synthesize cholesterol) responds to swelling but not temperature ( 91 ), whereas the mammalian channel is optimally active at ∼34–28°C.…”

Section: Discussionsupporting

confidence: 90%

“…Allosteric sites may include Cholesterol Recognition/Interaction Amino acid Consensus (CRAC)-like (KDLFRFLL) recognition motifs that span Loop 4 -TM5 (28,82,84). Consistent with our results, loss of TRPV4-cholesterol interaction in the TRPV4 R616Q CARC mutation was associated with gain-offunction for TRPV4 (67). Taken together, evidence suggests that cholesterol influences the context of the cellular sensory response such that TRPV4 activity in yeast (which cannot synthesize cholesterol) responds to swelling but not temperature (79), whereas the mammalian channel is J o u r n a l P r e -p r o o f optimally active at ~34-28 o C. Such context-dependence was observed in the TRPV1 channel, in which MβCD inhibits capsaicin-and proton-evoked currents (80,81) while facilitating its sensitivity to thermal inputs (80).…”

Section: Discussionsupporting

confidence: 88%

“…The outflow of aqueous humor is fine tuned by arrays of mechanosensitive molecules that include lipid rafts, caveolae, focal complexes, changes in gene expression, ECM and TRPV4, Piezo1, and TREK-1 channels ( 26 , 33 , 98 , 99 , 100 , 101 , 102 , 103 ). TRPV4 in endothelial and smooth muscle cells was reported to interact with Cav-1 ( 31 , 32 , 90 ), and both proteins regulate TM mechanosensitivity and outflow homeostasis ( 26 , 27 , 104 , 105 , 106 ). However, our hypothesis that TM cells manifest TRPV4-Cav-1 interactions in cholesterol-enriched raft/caveolar domains was not confirmed.…”

Section: Discussionmentioning

confidence: 99%

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Membrane cholesterol regulates TRPV4 function, cytoskeletal expression, and the cellular response to tension

Lakk

Hoffmann

Gorusupudi

et al. 2021

Journal of Lipid Research

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Despite the association of cholesterol with debilitating pressure-related diseases such as glaucoma, heart disease, and diabetes, its role in mechanotransduction is not well understood. We investigated the relationship between mechanical strain, free membrane cholesterol, actin cytoskeleton, and the stretch-activated TRPV4 (transient receptor potential vanilloid isoform 4) channel in human trabecular meshwork (TM) cells. Physiological levels of cyclic stretch resulted in time-dependent decreases in membrane cholesterol/phosphatidylcholine ratio and upregulation of stress fibers. Depleting free membrane cholesterol with m--cyclodextrin (MβCD) augmented TRPV4 activation by the agonist GSK1016790A, swelling and strain, with the effects reversed by cholesterol supplementation. MβCD increased membrane expression of TRPV4, caveolin-1, and flotillin. TRPV4 did not colocalize or interact with caveolae or lipid rafts, apart from a truncated ~75 kDa variant partially precipitated by a caveolin-1 antibody. MβCD induced currents in TRPV4-expressing Xenopus laevis oocytes. Thus, membrane cholesterol regulates trabecular transduction of mechanical information, with TRPV4 channels mainly located outside of cholesterol-enriched membrane domains. Moreover, the biomechanical milieu itself shapes the lipid content of TM membranes. Diet, cholesterol metabolism, and mechanical stress might modulate the conventional outflow pathway and intraocular pressure in glaucoma and diabetes.

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Ratio of Hydrophobic–Hydrophilic and Positive–Negative Residues at Lipid–Water-Interface Influences Surface Expression and Channel Gating of TRPV1

et al. 2022

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During evolution, TRPV1 has lost, retained or selected certain residues at Lipid-Water-Interface (LWI) and formed specific patterns there. The ratio of "hydrophobic-hydrophilic" and "positive-negative charged" residues at the inner LWI remains conserved throughout vertebrate evolution and play important role in regulating TRPV1 trafficking, localization and functions. Arg575 is an important residue as Arg575Asp mutant has reduced Capsaicin-sensitivity, surface expression, colocalization with lipid-raft markers, cell area, and increased cell lethality. This lethality is due to the disruption of the ratio between positive-negative charges there. Such lethality can be rescued by either using TRPV1-specfic inhibitor 5'-IRTX or by restoring the positive-negative charge ratio at that position, i.e. by introducing Asp576Arg mutation in Arg575Asp backbone. We propose that Arg575Asp mutant confers TRPV1 in a "constitutive-open-like" condition. These findings have broader implication in understanding the molecular basis of thermo-gating and channel-gating and the microenvironments involved in such process that goes erratic in different diseases.

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TRPV4 expresses in bone cell lineages and TRPV4-R616Q mutant causing Brachyolmia in human reveals “loss-of-interaction” with cholesterol

Cited by 23 publications

References 19 publications

Membrane cholesterol regulates TRPV4 function, cytoskeletal expression, and the cellular response to tension

Membrane cholesterol regulates TRPV4 function, cytoskeletal expression, and the cellular response to tension

Membrane cholesterol regulates TRPV4 function, cytoskeletal expression, and the cellular response to tension

Ratio of Hydrophobic–Hydrophilic and Positive–Negative Residues at Lipid–Water-Interface Influences Surface Expression and Channel Gating of TRPV1

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