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Mechanotransduction, the conversion of mechanical stimuli into electrical signals, is a fundamental process underlying essential physiological functions such as touch and pain sensing, hearing, and proprioception. Although the mechanisms for some of these functions have been identified, the molecules essential to the sense of pain have remained elusive. Here we report identification of TACAN (Tmem120A), an ion channel involved in sensing mechanical pain. TACAN is expressed in a subset of nociceptors, and its heterologous expression increases mechanically evoked currents in cell lines. Purification and reconstitution of TACAN in synthetic lipids generates a functional ion channel. Finally, a nociceptor-specific inducible knockout of TACAN decreases the mechanosensitivity of nociceptors and reduces behavioral responses to painful mechanical stimuli but not to thermal or touch stimuli. We propose that TACAN is an ion channel that contributes to sensing mechanical pain.
Assuntos
Canais Iônicos/fisiologia , Mecanotransdução Celular/genética , Nociceptores/metabolismo , Dor/genética , Tato/genética , Animais , Regulação da Expressão Gênica/genética , Humanos , Canais Iônicos/genética , Lipídeos/genética , Camundongos , Camundongos Knockout , Dor/fisiopatologia , Técnicas de Patch-Clamp , Estresse Mecânico , Tato/fisiologiaRESUMO
Biological membranes have a stunning ability to adapt their composition in response to physiological stress and metabolic challenges. Little is known how such perturbations affect individual organelles in eukaryotic cells. Pioneering work has provided insights into the subcellular distribution of lipids in the yeast Saccharomyces cerevisiae, but the composition of the endoplasmic reticulum (ER) membrane, which also crucially regulates lipid metabolism and the unfolded protein response, remains insufficiently characterized. Here, we describe a method for purifying organelle membranes from yeast, MemPrep. We demonstrate the purity of our ER membrane preparations by proteomics, and document the general utility of MemPrep by isolating vacuolar membranes. Quantitative lipidomics establishes the lipid composition of the ER and the vacuolar membrane. Our findings provide a baseline for studying membrane protein biogenesis and have important implications for understanding the role of lipids in regulating the unfolded protein response (UPR). The combined preparative and analytical MemPrep approach uncovers dynamic remodeling of ER membranes in stressed cells and establishes distinct molecular fingerprints of lipid bilayer stress.
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Bicamadas Lipídicas , Proteínas de Saccharomyces cerevisiae , Bicamadas Lipídicas/metabolismo , Saccharomyces cerevisiae/metabolismo , Estresse do Retículo Endoplasmático/fisiologia , Proteínas de Saccharomyces cerevisiae/metabolismo , Resposta a Proteínas não Dobradas , Retículo Endoplasmático/metabolismo , Tecnologia , Metabolismo dos LipídeosRESUMO
Different mechanisms driving a linear temperature dependence of the resistivity ρ â¼ T at van Hove singularities (VHSs) or metal-insulator transitions when doping a Mott insulator are being debated intensively with competing theoretical proposals. We experimentally investigate this using the exceptional tunability of twisted bilayer (TB) WSe2 by tracking the parameter regions where linear-in-T resistivity is found in dependency of displacement fields, filling, and magnetic fields. We find that even when the VHSs are tuned rather far away from the half-filling point and the Mott insulating transition is absent, the T-linear resistivity persists at the VHSs. When doping away from the VHSs, the T-linear behavior quickly transitions into a Fermi liquid behavior with a T2 relation. No apparent dependency of the linear-in-T resistivity, besides a rather strong change of prefactor, is found when applying displacement fields as long as the filling is tuned to the VHSs, including D â¼ 0.28 V/nm where a high-order VHS is expected. Intriguingly, such non-Fermi liquid linear-in-T resistivity persists even when magnetic fields break the spin-degeneracy of the VHSs at which point two linear in T regions emerge, for each of the split VHSs separately. This points to a mechanism of enhanced scattering at generic VHSs rather than only at high-order VHSs or by a quantum critical point during a Mott transition. Our findings provide insights into the many-body consequences arising out of VHSs, especially the non-Fermi liquid behavior found in moiré materials.
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Lipid droplets (LDs) are the main organelles for lipid storage, and their surfaces contain unique proteins with diverse functions, including those that facilitate the deposition and mobilization of LD lipids. Among organelles, LDs have an unusual structure with an organic, hydrophobic oil phase covered by a phospholipid monolayer. The unique properties of LD monolayer surfaces require proteins to localize to LDs by distinct mechanisms. Here we review the two pathways known to mediate direct LD protein localization: the CYTOLD pathway mediates protein targeting from the cytosol toLDs, and the ERTOLD pathway functions in protein targeting from the endoplasmic reticulum toLDs. We describe the emerging principles for each targeting pathway in animal cells and highlight open questions in the field.
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Retículo Endoplasmático , Gotículas Lipídicas , Animais , Citosol/metabolismo , Retículo Endoplasmático/metabolismo , Gotículas Lipídicas/metabolismo , Metabolismo dos Lipídeos , Transporte Proteico , Proteínas/metabolismoRESUMO
Anisotropic hopping in a toy Hofstadter model was recently invoked to explain a rich and surprising Landau spectrum measured in twisted bilayer graphene away from the magic angle. Suspecting that such anisotropy could arise from unintended uniaxial strain, we extend the Bistritzer-MacDonald model to include uniaxial heterostrain and present a detailed analysis of its impact on band structure and magnetotransport. We find that such strain strongly influences band structure, shifting the three otherwise-degenerate van Hove points to different energies. Coupled to a Boltzmann magnetotransport calculation, this reproduces previously unexplained nonsaturating [Formula: see text] magnetoresistance over broad ranges of density near filling [Formula: see text] and predicts subtler features that had not been noticed in the experimental data. In contrast to these distinctive signatures in longitudinal resistivity, the Hall coefficient is barely influenced by strain, to the extent that it still shows a single sign change on each side of the charge neutrality point-surprisingly, this sign change no longer occurs at a van Hove point. The theory also predicts a marked rotation of the electrical transport principal axes as a function of filling even for fixed strain and for rigid bands. More careful examination of interaction-induced nematic order versus strain effects in twisted bilayer graphene could thus be in order.
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Mechanical forces modify the cell membrane potential by opening mechanosensitive ion channels. We report the design and construction of a lipid bilayer tensiometer to study channels that respond to lateral membrane tension, [Formula: see text]â, in the range 0.2 to 1.4 [Formula: see text] (0.8 to 5.7 [Formula: see text]â). The instrument consists of a black-lipid-membrane bilayer, a custom-built microscope, and a high-resolution manometer. Values of [Formula: see text] are obtained from the determination of the bilayer curvature as a function of applied pressure by means of the Young-Laplace equation. We demonstrate that [Formula: see text] can be determined by calculating the bilayer radius of curvature from fluorescence microscopy imaging or from measurements of the bilayer's electrical capacitance, both yielding similar results. Using electrical capacitance, we show that the mechanosensitive potassium channel TRAAK responds to [Formula: see text]â, not curvature. TRAAK channel open probability increases as [Formula: see text] is increased from 0.2 to 1.4 [Formula: see text] but open probability never reaches 0.5. Thus, TRAAK opens over a wide range of [Formula: see text]â, but with a tension sensitivity about one-fifth that of the bacterial mechanosensitive channel MscL.
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Canais Iônicos , Bicamadas Lipídicas , Canais Iônicos/metabolismo , Bicamadas Lipídicas/metabolismo , Canais de Potássio/metabolismoRESUMO
The synthesis and characterization of small boron clusters with unique size and regular arrangement are crucial for boron chemistry and two-dimensional borophene materials. In this study, together with theoretical calculations, the joint molecular beam epitaxy and scanning tunneling microscopy experiments achieve the formation of unique B5 clusters on monolayer borophene (MLB) on a Cu(111) surface. The B5 clusters tend to selectively bind to specific sites of MLB with covalent boron-boron bonds in the periodic arrangement, which can be ascribed to the charge distribution and electron delocalization character of MLB and also prohibits nearby co-adsorption of B5 clusters. Furthermore, the close-packed adsorption of B5 clusters would facilitate the synthesis of bilayer borophene, exhibiting domino effect-like growth mode. The successful growth and characterization of uniform boron clusters on a surface enrich the boron-based nanomaterials and reveal the essential role of small clusters during the growth of borophene.
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Bax proteins form pores in the mitochondrial outer membrane to initiate apoptosis. This might involve their embedding in the cytosolic leaflet of the lipid bilayer, thus generating tension to induce a lipid pore with radially arranged lipids forming the wall. Alternatively, Bax proteins might comprise part of the pore wall. However, there is no unambiguous structural evidence for either hypothesis. Using NMR, we determined a high-resolution structure of the Bax core region, revealing a dimer with the nonpolar surface covering the lipid bilayer edge and the polar surface exposed to water. The dimer tilts from the bilayer normal, not only maximizing nonpolar interactions with lipid tails but also creating polar interactions between charged residues and lipid heads. Structure-guided mutations demonstrate the importance of both types of protein-lipid interactions in Bax pore assembly and core dimer configuration. Therefore, the Bax core dimer forms part of the proteolipid pore wall to permeabilize mitochondria.
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Mitocôndrias/metabolismo , Membranas Mitocondriais/metabolismo , Proteína X Associada a bcl-2/metabolismo , Apoptose/fisiologia , Humanos , Bicamadas Lipídicas/metabolismoRESUMO
The unfolded protein response (UPR) is a conserved homeostatic program that is activated by misfolded proteins in the lumen of the endoplasmic reticulum (ER). Recently, it became evident that aberrant lipid compositions of the ER membrane, referred to as lipid bilayer stress, are equally potent in activating the UPR. The underlying molecular mechanism, however, remained unclear. We show that the most conserved transducer of ER stress, Ire1, uses an amphipathic helix (AH) to sense membrane aberrancies and control UPR activity. In vivo and in vitro experiments, together with molecular dynamics (MD) simulations, identify the physicochemical properties of the membrane environment that control Ire1 oligomerization. This work establishes the molecular mechanism of UPR activation by lipid bilayer stress.
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Estresse do Retículo Endoplasmático , Retículo Endoplasmático/metabolismo , Membranas Intracelulares/metabolismo , Bicamadas Lipídicas/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Resposta a Proteínas não Dobradas , Glicoproteínas de Membrana/química , Glicoproteínas de Membrana/genética , Simulação de Dinâmica Molecular , Mutação , Conformação Proteica em alfa-Hélice , Dobramento de Proteína , Proteínas Serina-Treonina Quinases/química , Proteínas Serina-Treonina Quinases/genética , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/genética , Transdução de Sinais , Relação Estrutura-Atividade , Fatores de TempoRESUMO
SignificanceWhen two sheets of graphene are twisted to the magic angle of 1.1∘, the resulting flat moiré bands can host exotic correlated electronic states such as superconductivity and ferromagnetism. Here, we show transport properties of a twisted bilayer graphene device at 1.38∘, far enough above the magic angle that we do not expect exotic correlated states. Instead, we see several unusual behaviors in the device's resistivity upon tuning both charge carrier density and perpendicular magnetic field. We can reproduce these behaviors with a surprisingly simple model based on Hofstadter's butterfly. These results shed light on the underlying properties of twisted bilayer graphene.
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Cell membranes are complex assemblies of proteins and lipids making transient or long-term associations that have yet to be characterized at a molecular level. Here, cryo-electron microscopy is applied to determine how phospholipids and cholesterol arrange between neighboring proteins (nicotinic acetylcholine receptors) of Torpedo cholinergic membrane. The lipids exhibit distinct properties in the two leaflets of the bilayer, influenced by the protein surfaces and by differences in cholesterol concentration. In the outer leaflet, the lipids show no consistent motif away from the protein surfaces, in keeping with their assumed fluidity. In the inner leaflet, where the cholesterol concentration is higher, the lipids organize into extensive close-packed linear arrays. These arrays are built from the sterol groups of cholesterol and the initial saturated portions of the phospholipid hydrocarbon chains. Together, they create an ordered â¼7 Å-thick "skin" within the hydrophobic core of the bilayer. The packing of lipids in the arrays appears to bear a close relationship to the linear cholesterol arrays that form crystalline monolayers at the air-water interface.
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Membrana Celular , Colesterol , Fosfolipídeos , Animais , Membrana Celular/ultraestrutura , Colesterol/metabolismo , Microscopia Crioeletrônica , Bicamadas Lipídicas , Fluidez de Membrana , Fosfolipídeos/metabolismo , TorpedoRESUMO
The 99-residue C-terminal domain of amyloid precursor protein (APP-C99), precursor to amyloid beta (Aß), is a transmembrane (TM) protein containing intrinsically disordered N- and C-terminal extramembrane domains. Using molecular dynamics (MD) simulations, we show that the structural ensemble of the C99 monomer is best described in terms of thousands of states. The C99 monomer has a propensity to form ß-strand in the C-terminal extramembrane domain, which explains the slow spin relaxation times observed in paramagnetic probe NMR experiments. Surprisingly, homodimerization of C99 not only narrows the conformational ensemble from thousands to a few states through the formation of metastable ß-strands in extramembrane domains but also stabilizes extramembrane α-helices. The extramembrane domain structure is observed to dramatically impact the homodimerization motif, resulting in the modification of TM domain conformations. Our study provides an atomic-level structural basis for communication between the extramembrane domains of the C99 protein and TM homodimer formation. This finding could serve as a general model for understanding the influence of disordered extramembrane domains on TM protein structure.
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Peptídeos beta-Amiloides , Precursor de Proteína beta-Amiloide , Precursor de Proteína beta-Amiloide/metabolismo , Dimerização , Peptídeos beta-Amiloides/metabolismo , Conformação Proteica em Folha beta , Domínios Proteicos , Secretases da Proteína Precursora do Amiloide/metabolismoRESUMO
Coordinated cell function requires a variety of subcellular organelles to exchange proteins and lipids across physical contacts that are also referred to as membrane contact sites. Such organelle-to-organelle contacts also evoke interest because they can appear in response to metabolic changes, immune activation, and possibly other stimuli. The microscopic size and complex, crowded geometry of these contacts, however, makes them difficult to visualize, manipulate, and understand inside cells. To address this shortcoming, we deposited endoplasmic reticulum (ER)-enriched microsomes purified from rat liver or from cultured cells on a coverslip in the form of a proteinaceous planar membrane. We visualized real-time lipid and protein exchange across contacts that form between this ER-mimicking membrane and lipid droplets (LDs) purified from the liver of rat. The high-throughput imaging possible in this geometry reveals that in vitro LD-ER contacts increase dramatically when the metabolic state is changed by feeding the animal and also when the immune system is activated. Contact formation in both cases requires Rab18 GTPase and phosphatidic acid, thus revealing common molecular targets operative in two very different biological pathways. An optical trap is used to demonstrate physical tethering of individual LDs to the ER-mimicking membrane and to estimate the strength of this tether. These methodologies can potentially be adapted to understand and target abnormal contact formation between different cellular organelles in the context of neurological and metabolic disorders or pathogen infection.
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Retículo Endoplasmático , Gotículas Lipídicas , Animais , Células Cultivadas , Retículo Endoplasmático/imunologia , Retículo Endoplasmático/metabolismo , Gotículas Lipídicas/imunologia , Gotículas Lipídicas/metabolismo , Metabolismo dos Lipídeos , Microssomos Hepáticos/química , Membranas Mitocondriais/metabolismo , Ácidos Fosfatídicos/metabolismo , Ratos , Proteínas rab de Ligação ao GTP/metabolismoRESUMO
We propose minimal transport experiments in the coherent regime that can probe the chirality of twisted moiré structures. We show that only with a third contact and in the presence of an in-plane magnetic field (or another time-reversal symmetry breaking effect) a chiral system may display nonreciprocal transport in the linear regime. We then propose to use the third lead as a voltage probe and show that opposite enantiomers give rise to different voltage drops on the third lead. Additionally, in the scenario of layer-discriminating contacts, the third lead can serve as a current probe capable of detecting different handedness even in the absence of a magnetic field. In a complementary configuration, applying opposite voltages on the two layers of the third lead gives rise to a chiral (super)current in the absence of a source-drain voltage whose direction is determined by its chirality.
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The superconducting coplanar waveguide (SCPW) cavity plays an essential role in various areas like superconducting qubits, parametric amplifiers, radiation detectors, and studying magnon-photon and photon-phonon coupling. Despite its wide-ranging applications, the use of SCPW cavities to study various van der Waals 2D materials has been relatively unexplored. The resonant modes of the SCPW cavity exquisitely sense the dielectric environment. In this work, we measure the charge compressibility of bilayer graphene coupled to a half-wavelength SCPW cavity. Our approach provides a means to detect subtle changes in the capacitance of the bilayer graphene heterostructure, which depends on the compressibility of bilayer graphene, manifesting as shifts in the resonant frequency of the cavity. This method holds promise for exploring a wide class of van der Waals 2D materials, including transition metal dichalcogenides (TMDs) and their moiré, where DC transport measurement is challenging.
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We present experimental findings on electron-electron scattering in two-dimensional moiré heterostructures with a tunable Fermi wave vector, reciprocal lattice vector, and band gap. We achieve this in high-mobility aligned heterostructures of bilayer graphene (BLG) and hBN. Around the half-full point, the primary contribution to the resistance of these devices arises from Umklapp electron-electron (Uee) scattering, making the resistance of graphene/hBN moiré devices significantly larger than that of non-aligned devices (where Uee is forbidden). We find that the strength of Uee scattering follows a universal scaling with Fermi energy and is nonmonotonically dependent on the superlattice period. The Uee scattering can be tuned with the electric field and is affected by layer polarization of BLG. It has a strong particle-hole asymmetry; the resistance when the chemical potential is in the conduction band is significantly lower than when it is in the valence band, making the electron-doped regime more practical for potential applications.
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At near-parallel orientation, twisted bilayers of transition metal dichalcogenides exhibit interlayer charge transfer-driven out-of-plane ferroelectricity. Here, we report detailed electrical transport in a dual-gated graphene field-effect transistor placed on a 2.1° twisted bilayer WSe2. We observe hysteretic transfer characteristics and an emergent charge inhomogeneity with multiple local Dirac points evolving with an increasing electric displacement field (D). Concomitantly, we also observe a strong nonlocal voltage signal at D â¼ 0 V/nm that decreases rapidly with increasing D. A linear scaling of the nonlocal signal with longitudinal resistance suggests edge mode transport, which we attribute to the breaking of valley symmetry of graphene due to the spatially fluctuating electric field from the underlying polarized moiré domains. A quantitative analysis suggests the emergence of finite-size domains in graphene that modulate the charge and the valley currents simultaneously. This work underlines the impact of interfacial ferroelectricity that can trigger a new generation of devices.
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We implement circuit quantum electrodynamics (cQED) with quantum dots in bilayer graphene, a maturing material platform that can host long-lived spin and valley states. Our device combines a high-impedance (Zr ≈ 1 kΩ) superconducting microwave resonator with a double quantum dot electrostatically defined in a graphene-based van der Waals heterostructure. Electric dipole coupling between the subsystems allows the resonator to sense the electric susceptibility of the double quantum dot from which we reconstruct its charge stability diagram. We achieve sensitive and fast detection of the interdot transition with a signal-to-noise ratio of 3.5 within 1 µs integration time. The charge-photon interaction is quantified in the dispersive and resonant regimes by comparing the resonator response to input-output theory, yielding a coupling strength of g/2π = 49.7 MHz. Our results introduce cQED as a probe for quantum dots in van der Waals materials and indicate a path toward coherent charge-photon coupling with bilayer graphene quantum dots.
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Van Hove singularities enhance many-body interactions and induce collective states of matter ranging from superconductivity to magnetism. In magic-angle twisted bilayer graphene, van Hove singularities appear at low energies and are malleable with density, leading to a sequence of Lifshitz transitions and resets observable in Hall measurements. However, without a magnetic field, linear transport measurements have limited sensitivity to the band's topology. Here, we utilize nonlinear longitudinal and transverse transport measurements to probe these unique features in twisted bilayer graphene at zero magnetic field. We demonstrate that the nonlinear responses, induced by the Berry curvature dipole and extrinsic scattering processes, intricately map the Fermi surface reconstructions at various fillings. Importantly, our experiments highlight the intrinsic connection of these features with the moiré bands. Beyond corroborating the insights from linear Hall measurements, our findings establish nonlinear transport as a pivotal tool for probing band topology and correlated phenomena.
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Double-layer quantum systems are promising platforms for realizing novel quantum phases. Here, we report a study of quantum oscillations (QOs) in a weakly coupled double-layer system composed of a large-angle twisted-double-bilayer graphene (TDBG). We quantify the interlayer coupling strength by measuring the interlayer capacitance from the QOs pattern at low temperatures, revealing electron-hole asymmetry. At high temperatures when SdHOs are thermally smeared, we observe resistance peaks when Landau levels (LLs) from two moiré minivalleys are aligned, regardless of carrier density; eventually, it results in a 2-fold increase of oscillating frequency in D, serving as compelling evidence of the magneto-intersub-band oscillations (MISOs) in double-layer systems. The temperature dependence of MISOs suggests that electron-electron interactions play a crucial role and the scattering times obtained from MISO thermal damping are correlated with the interlayer coupling strength. Our study reveals intriguing interplays among Landau quantization, moiré band structure, and scatterings.