Crosstalk between regulatory elements in disordered TRPV4 N-terminus modulates lipid-dependent channel activity

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: July 13, 2023

Abstract Intrinsically disordered regions (IDRs) are essential for membrane receptor regulation but often remain unresolved in structural studies. TRPV4, a member of the TRP vanilloid channel family involved thermo- and osmosensation, has large N-terminal IDR approximately 150 amino acids. With an integrated biology approach, we analyze ensemble TRPV4 network antagonistic regulatory elements it encodes. These modulate activity hierarchical lipid-dependent manner through transient long-range interactions. A highly conserved autoinhibitory patch acts as master regulator by competing with PIP 2 binding to attenuate activity. Molecular dynamics simulations show that loss interaction between -binding site reduces force exerted on structured core TRPV4. This work demonstrates coupled highlights importance IDRs function regulation.

Language: Английский

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Membranes under the Magnetic Lens: A Dive into the Diverse World of Membrane Protein Structures Using Cryo-EM

Chemical Reviews, Journal Year: 2022, Volume and Issue: 122(17), P. 13989 - 14017

Published: July 18, 2022

Membrane proteins are highly diverse in both structure and function can, therefore, present different challenges for determination. They biologically important cells organisms as gatekeepers information molecule transfer across membranes, but each class of membrane can unique obstacles to Historically, many protein structures have been investigated using engineered constructs or larger fusion improve solubility and/or increase particle size. Other strategies included the deconstruction full-length target smaller soluble domains. These manipulations were often required crystal formation support X-ray crystallography circumvent lower resolution due high noise dynamic motions subdomains. However, recent revolutions biochemistry cryo-electron microscopy now provide an opportunity solve large, >1 megadalton (MDa), small, <100 kDa (kDa), drug targets near-native conditions, routinely reaching resolutions around below 3 Å. This review provides insights into how advances biology biochemistry, well technical microscopy, help us a large variety groups, from small receptors transporters more complex machineries.

Language: Английский

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Monitoring the conformational ensemble and lipid environment of a mechanosensitive channel under cyclodextrin-induced membrane tension

Structure, Journal Year: 2024, Volume and Issue: 32(6), P. 739 - 750.e4

Published: March 22, 2024

Membrane forces shift the equilibria of mechanosensitive channels enabling them to convert mechanical cues into electrical signals. Molecular tools stabilize and methods capture their highly dynamic states are lacking. Cyclodextrins can mimic tension through sequestering lipids from membranes. Here we probe conformational ensemble MscS by EPR spectroscopy, lipid environment with NMR, function electrophysiology under cyclodextrin-induced tension. We show extent activation depends on cyclodextrin-to-lipid ratio, that depleted slower when is present. This has implications in MscS' kinetics distinct membrane scaffolds such as nanodiscs or liposomes used. find transits closed sub-conducting state(s) before it desensitizes, due lack availability its vicinity required for closure. Our approach allows monitoring tension-sensitive proteins screening molecules capable inducing molecular bilayers.

Language: Английский

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Analysis of hyperforin (St. John’s wort) action at TRPC6 channel leads to the development of a new class of antidepressant drugs

Molecular Psychiatry, Journal Year: 2022, Volume and Issue: 27(12), P. 5070 - 5085

Published: Oct. 12, 2022

Abstract St. John’s wort is an herb, long used in folk medicine for the treatment of mild depression. Its antidepressant constituent, hyperforin, has properties such as chemical instability and induction drug-drug interactions that preclude its use individual pharmacotherapies. Here we identify transient receptor potential canonical 6 channel (TRPC6) a druggable target to control anxious depressive behavior requirement hyperforin action. We demonstrate TRPC6 deficiency mice not only results behavior, but also reduces excitability hippocampal CA1 pyramidal neurons dentate gyrus granule cells. Using electrophysiology targeted mutagenesis, show activates via specific binding motif at TRPC6. performed analysis action develop new drug uses same mechanism synthesized analog Hyp13, which shows similar recapitulates TRPC6-dependent anxiolytic effects mice. Hyp13 does activate pregnan-X-receptor ( PXR ) thereby loses induce interactions. This may provide approach better treatments depression, since depression remains one most treatment-resistant mental disorders, warranting development effective drugs based on naturally occurring compounds.

Language: Английский

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Cannabinoid non-cannabidiol site modulation of TRPV2 structure and function

Nature Communications, Journal Year: 2022, Volume and Issue: 13(1)

Published: Dec. 5, 2022

Abstract TRPV2 is a ligand-operated temperature sensor with poorly defined pharmacology. Here, we combine calcium imaging and patch-clamp electrophysiology cryo-electron microscopy (cryo-EM) to explore how activity modulated by the phytocannabinoid Δ 9 -tetrahydrocannabiorcol (C16) probenecid. C16 probenecid act in concert stimulate responses including histamine release from rat human mast cells. Each ligand causes distinct conformational changes as revealed cryo-EM. Although binding for remains elusive, associates within vanilloid pocket. As such, location that of cannabidiol, partially overlapping site inhibitor piperlongumine. Taken together, discover new cannabinoid under influence allosteric control This molecular insight into modulation enhances our understanding normal pathophysiology.

Language: Английский

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Electrostatic switch mechanisms of membrane protein trafficking and regulation

Biophysical Reviews, Journal Year: 2023, Volume and Issue: 15(6), P. 1967 - 1985

Published: Dec. 1, 2023

Abstract Lipid-protein interactions are normally classified as either specific or general. Specific refer to lipid binding sites within a membrane protein, thereby modulating the protein’s thermal stability kinetics. General indirect effects whereby lipids affect proteins by membrane’s physical properties, e.g., its fluidity, thickness, dipole potential. It is not widely recognized that there third distinct type of lipid-protein interaction. Intrinsically disordered N- C-termini can interact directly but nonspecifically with surrounding membrane. Many peripheral held cytoplasmic surface plasma via cooperative combination two forces: hydrophobic anchoring and electrostatic attraction. An acyl chain, myristoyl, added post-translationally one termini inserts itself into matrix helps hold onto Electrostatic attraction occurs between positively charged basic amino acid residues (lysine arginine) on terminal tails negatively phospholipid head groups, such phosphatidylserine. Phosphorylation serine tyrosine regulatory protein kinases allows for an switch mechanism control trafficking protein. Kinase action reduces positive charge tail, weakening releasing from A similar regulates many integral proteins, here only involved, modulates activity altering stabilities different conformational states.

Language: Английский

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Harmonizing Magnetic Mitohormetic Regenerative Strategies: Developmental Implications of a Calcium–Mitochondrial Axis Invoked by Magnetic Field Exposure

Bioengineering, Journal Year: 2023, Volume and Issue: 10(10), P. 1176 - 1176

Published: Oct. 10, 2023

Mitohormesis is a process whereby mitochondrial stress responses, mediated by reactive oxygen species (ROS), act cumulatively to either instill survival adaptations (low ROS levels) or produce cell damage (high levels). The mitohormetic nature of extremely low-frequency electromagnetic field (ELF-EMF) exposure thus makes it susceptible extraneous influences that also impinge on production and contribute the collective response. Consequently, magnetic stimulation paradigms are prone experimental variability depending diverse circumstances. failure, inability, control for these factors has contributed existing discrepancies between published reports in interpretations made from results generated therein. Confounding environmental include ambient fields, temperature, mechanical environment, conventional use aminoglycoside antibiotics. Biological type seeding density as well developmental, inflammatory, senescence statuses cells depend prior handling sample. Technological aspects directionality, uniformity, amplitude, duration exposure. All will exhibit manifestations at level culminate unified cellular response conjunction with Fortunately, many under experimenter. This review focus delineating areas requiring technical biological harmonization assist designing therapeutic strategies more clearly defined better predicted outcomes improve mechanistic interpretation data, rather than precise applications. explore underlying similarities other forms biophysical stimuli, such mutually induce elevations intracellular calcium prerequisite outcome. These stimuli commonly invoke activity transient receptor potential cation channel classes, TRPC1.

Language: Английский

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Stay connected: The myoendothelial junction proteins in vascular function and dysfunction

Vascular Pharmacology, Journal Year: 2025, Volume and Issue: 158, P. 107463 - 107463

Published: Jan. 14, 2025

Language: Английский

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Pharmacology of TRPC Channels and Its Potential in Cardiovascular and Metabolic Medicine

The Annual Review of Pharmacology and Toxicology, Journal Year: 2021, Volume and Issue: 62(1), P. 427 - 446

Published: Sept. 9, 2021

Transient receptor potential canonical (TRPC) proteins assemble to form homo- or heterotetrameric, nonselective cation channels permeable K+, Na+, and Ca2+. TRPC are thought act as complex integrators of physical chemical environmental stimuli. Although the understanding essential physiological roles is incomplete, their implication in various pathological mechanisms conditions nervous system, kidneys, cardiovascular system combination with lack major adverse effects knockout channel inhibition driving search modulators therapeutics. Here, we review most promising small-molecule modulators, mode action, study treatment metabolic disease.

Language: Английский

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Phosphorylation of distal C-terminal residues promotes TRPV4 channel activation in response to arachidonic acid

Journal of Biological Chemistry, Journal Year: 2025, Volume and Issue: unknown, P. 108260 - 108260

Published: Feb. 1, 2025

Transient receptor potential vanilloid 4 (TRPV4) is a Ca2+-permeable channel activated by diverse physical and chemical stimuli, including mechanical stress endogenous lipid arachidonic acid (AA) its metabolites. Phosphorylation of TRPV4 protein kinase A (PKA) C (PKC) predominant mechanism for regulation, especially in the cytoplasmic domains due to their importance assembly, channelopathies. However, studies corresponding phosphorylation sites these kinases remain incomplete. We investigated role Ser-823 residue as site regulating activity agonist-induced activation. Using mass spectrometry, we identified new confirmed previously known Ser-824 C-terminal tail. The low level at was stimulated PKC lesser extent PKA human coronary artery endothelial cells (HCAECs) embryonic kidney 293 (HEK 293) cells. AA-induced activation enhanced phosphomimetic S823E but blunted S823A/S824A mutants, whereas synthetic agonist GSK1016790A unaffected. Further, AA not or abolished inhibitor alone combination with inhibitor, respectively. computational modeling, refined proposed structural model regulation phosphorylation. Together, results provide insight into how stimuli-specific regulated discrete residues (e.g., Ser-824) channel.

Language: Английский

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