A single N-terminal cysteine in TRPV1 determines activation by pungent compounds from onion and garlic

Salazar, Héctor; Llorente, Itzel; Jara-Oseguera, Andrés; García-Villegas, Refugio; Munari, Mika; Gordon, Sharona E.; Islas, León D; Rosenbaum, Tamara

doi:10.1038/nn2056

Cited by 196 publications

(191 citation statements)

References 36 publications

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“…An agonist with the potential to stimulate activity-dependent therapeutic molecule transport will be most useful if it also exhibits a parallel increase of potency or efficacy for sensitized versus normal TRPV1. Protein kinase C activation (23)(24)(25)(26) and oxidative modification (20,27,28) are major biochemical pathways that enhance TRPV1 sensitivity to chemical agonists. PDBu and PAO, chemical activators stimulating PKC or mimicking cellular oxidation, respectively, could elicit TRPV1-dependent intracellular Ca 2+ rise (20,29).…”

Section: Resultsmentioning

confidence: 99%

Activity-dependent targeting of TRPV1 with a pore-permeating capsaicin analog

Wang

Chuang³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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The capsaicin receptor TRPV1 is the principal transduction channel for nociception. Excessive TRPV1 activation causes pathological pain. Ideal pain mangement requires selective inhibition of hyperactive pain-sensing neurons, but sparing normal nociception. We sought to determine whether it is possible to use activity-dependent TRPV1 agonists to identify nerves with excessive TRPV1 activity, as well as exploit the TRPV1 pore to deliver charged anesthetics for neuronal silencing. We synthesized a series of permanently charged capsaicinoids and found that one, cap-ET, efficaciously evoked TRPV1-dependent entry of Ca 2+ or the large cationic dye YO-PRO-1 comparably to capsaicin, but far smaller electrical currents. Cap-ETinduced YO-PRO-1 transport required permeation of both the agonist and the dye through the TRPV1 pore and could be enhanced by kinase activation or oxidative covalent modification. Moreover, cap-ET reduced capsaicin-induced currents by a voltage-dependent block of the pore. A low dose of cap-ET elicited entry of permanently charged Na + channel blockers to effectively suppress Na + currents in sensory neurons presensitized with oxidative chemicals. These results implicate therapeutic potential of these unique TRPV1 agonists exhibiting activity-dependent ion transport but of minimal pain-producing risks.activity-dependent capsaicinoids | hyperalgesia | ion permeation | selective analgesia C apsaicin, a small lipophilic molecule from hot chili peppers, acts on sensory neurons by opening the TRPV1 channel. TRPV1 is activated by noxious temperatures (>43°C) and serves as an integrator for major pain-producing signals (1, 2). Inflammatory hyperalgesia is dramatically reduced in mice lacking TRPV1 (3, 4). Besides being an attractive target for pain management, TRPV1 regulates autonomic function, such as body temperature, blood vessel tone, and release of transmitters from sensory nerves (5-9). TRPV1 activated by capsaicin at a concentration far below the pain-producing threshold triggers neuropeptide release (7) and production of other second messengers (10). In light of the complexity of TRPV1 actions in physiology, one major challenge in developing TRPV1-based pain treatment is to target therapeutic compounds to selectively inhibit hyperactive nociceptive neurons while sparing nerves of normal thresholds for pain detection.Capsaicin is among the most powerful chemical agonists of TRPV1. Being small and hydrophobic, capsaicin crosses the plasma membrane readily to reach its intracellular ligand-binding site on TRPV1 (11, 12), leading to channel activation and cation permeation. TRPV1 activation rapidly depolarizes nerves to evoke acute pain sensation and allows Ca 2+ entry to initiate downstream signaling events such as neuropeptide release or production of other second messengers. Nevertheless, TRPV1 activation facilitates cellular transport of organic cations such as tetraethylammonium (TEA), N-methylglucamine (NMG) (13), and the quaternary ammonium Na + channel blocker QX-314 (14) or even larger fluor...

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Section: Resultsmentioning

confidence: 99%

Activity-dependent targeting of TRPV1 with a pore-permeating capsaicin analog

Wang

Chuang³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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“…Various reactive chemicals such as mustard oil, cinnamaldehyde, and allicin (garlic) activate TRPA1 through cysteine modification, and 3 cytoplasmic cysteines juxtaposing the transmembrane domain in the N terminus of TRPA1 are involved in these responses. Interestingly, allicin was recently shown to activate TRPV1 through a cysteine located at the N-terminal region (35). Therefore, binding of chemicals to this linker region of TRP channels between ankyrin domains and transmembrane helices might cause conformational change and downstream ion channel gating.…”

Section: Discussionmentioning

confidence: 99%

Two amino acid residues determine 2-APB sensitivity of the ion channels TRPV3 and TRPV4

Grandl

Bandell

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

105

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Temperature-activated transient receptor potential ion channels (thermoTRPs) are polymodal detectors of various stimuli including temperature, voltage, and chemicals. To date, it is not known how TRP channels integrate the action of such disparate stimuli. Identifying specific residues required for channel-activation by distinct stimuli is necessary for understanding overall TRP channel function. TRPV3 is activated by warm temperatures and various chemicals, and is modulated by voltage. One potent activator of TRPV3 is 2-aminoethyl diphenylborinate (2-APB), a synthetic chemical that modulates many TRP channels. In a high-throughput mutagenesis screen of Ϸ14,000 mutated mouse TRPV3 clones, we found 2 residues (H426 and R696) specifically required for sensitivity of TRPV3 to 2-APB, but not to camphor or voltage. The cytoplasmic N-terminal mutation H426N in human, dog, and frog TRPV3 also effectively abolished 2-APB activation without affecting camphor responses. Interestingly, chicken TRPV3 is weakly sensitive to 2-APB, and the equivalent residue at 426 is an asparagine (N). Mutating this residue to histidine induced 2-APB sensitivity of chicken TRPV3 to levels comparable for other TRPV3 orthologs. The cytoplasmic C-terminal mutation R696K in the TRP box displayed 2-APB specific deficits only in the presence of extracellular calcium, suggesting involvement in gating. TRPV4, a related thermoTRP, is 2-APB insensitive and has variant sequences at both residues identified here. Remarkably, mutating these 2 residues in TRPV4 to TRPV3 sequences (N426H and W737R) was sufficient to induce TRPV3-like 2-APB sensitivity. Therefore, 2-APB activation of TRPV3 is separable from other activation mechanisms, and depends on 2 cytoplasmic residues.A subset of transient receptor potential (TRP) ion channels have important roles in sensory biology, including pain and thermosensation (1-4). The requirement of several so-called thermoTRPs in thermosensation in vivo has been demonstrated, suggesting that these ion channels have a physiological role as molecular thermometers (5-9). Thresholds of temperatureactivation of thermoTRPs are shifted in the presence of chemical agonists, effectively modulating thermosensation (1, 10-12). Several natural and synthetic compounds have been identified as agonists or antagonists for each of the thermoTRPs (13). However, little is known about how these chemicals modulate TRP channel activity.TRPV3, one of these thermoTRPs, is a nonselective cation channel expressed primarily in mammalian keratinocytes, and is involved in thermosensation (4,7,14). Like other thermoTRP channels, TRPV3 is a polymodal detector of temperature (heat) and chemicals. TRPV3 agonists include structurally-related natural compounds such as camphor, thymol, and carvacrol (15, 16). One of the most potent TRPV3 agonists is 2-aminoethyl diphenylborinate (2-APB), a synthetic compound that modulates the activity of many ion channels. It is a common activator of the heat-gated TRPV ion-channels TRPV1, TRPV2, and TRPV3 (17,18). Besides i...

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“…HEK293 Cell Culture and Capsaicin-induced Currents-HEK293 cells expressing large T antigen were transfected with wild-type and mutant pCDNA3-rTRPV1 and pIRES-GFP (BD Biosciences) with Lipofectamine (Invitrogen) following previously described methods (22,23). Inside-out and outside-out patch clamp recordings of TRPV1 were made using Ca 2ϩ -free symmetrical solutions consisting of 130 mM NaCl, 3 mM HEPES (pH 7.2), and 1 mM EDTA.…”

Section: Methodsmentioning

confidence: 99%

“…Pipettes were pulled from borosilicate glass, covered with dental wax to reduce stray capacitance, and fire polished to a resistance of 10 -15 M⍀ (22,23). Currents were filtered at 2 kHz and sampled at 5 kHz.…”

Section: Methodsmentioning

confidence: 99%

Identification of a Binding Motif in the S5 Helix That Confers Cholesterol Sensitivity to the TRPV1 Ion Channel

Picazo-Juárez

Romero-Suárez

Nieto-Posadas

et al. 2011

Journal of Biological Chemistry

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131

113

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The TRPV1 ion channel serves as an integrator of noxious stimuli with its activation linked to pain and neurogenic inflammation. Cholesterol, a major component of cell membranes, modifies the function of several types of ion channels. Here, using measurements of capsaicin-activated currents in excised patches from TRPV1-expressing HEK cells, we show that enrichment with cholesterol, but not its diastereoisomer epicholesterol, markedly decreased wild-type rat TRPV1 currents. Substitutions in the S5 helix, rTRPV1-R579D, and rTRPV1-F582Q, decreased this cholesterol response and rTRPV1-L585I was insensitive to cholesterol addition. Two human TRPV1 variants, with different amino acids at position 585, had different responses to cholesterol with hTRPV1-Ile 585 being insensitive to this molecule. However, hTRPV1-I585L was inhibited by cholesterol addition similar to rTRPV1 with the same S5 sequence. In the absence of capsaicin, cholesterol enrichment also inhibited TRPV1 currents induced by elevated temperature and voltage. These data suggest that there is a cholesterol-binding site in TRPV1 and that the functions of TRPV1 depend on the genetic variant and membrane cholesterol content. The transient receptor potential (TRP)3 family of ion channels is found throughout the animal kingdom and has been shown to subserve numerous functions. One extensively studied member of this family is the TRPV1 (Vanilloid 1) channel. Structurally, TRPV1 is thought to be a tetramer comprised of subunits each with six transmembrane domains (S1-S6), with the putative pore of the channel located between S5 and S6. It also contains large intracellular amino and carboxyl termini that have been shown to be involved both in channel gating and regulation (for review, see Ref.

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A single N-terminal cysteine in TRPV1 determines activation by pungent compounds from onion and garlic

Cited by 196 publications

References 36 publications

Activity-dependent targeting of TRPV1 with a pore-permeating capsaicin analog

Activity-dependent targeting of TRPV1 with a pore-permeating capsaicin analog

Two amino acid residues determine 2-APB sensitivity of the ion channels TRPV3 and TRPV4

Identification of a Binding Motif in the S5 Helix That Confers Cholesterol Sensitivity to the TRPV1 Ion Channel

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