Zinc activates damage-sensing TRPA1 ion channels

Hu, Hongzhen; Bandell, Michael; Petrus, Matt; Zhu, Michael X.; Patapoutian, Ardem

doi:10.1038/nchembio.146

Cited by 200 publications

(197 citation statements)

References 49 publications

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“…Addition of Zn 2 + had a similar effect to PP2 on the responses of TRPM8 variants to agonist, with the most noticeable enhancing effects on K423N and R485W. Rescue of TRPM8 mutants by Zn 2 + is a novel observation, but with similarities to the effects of Zn 2 + on mutated TRPA1 and suggestive of stabilization of miss-folded protein [10].…”

Section: Discussionmentioning

confidence: 83%

“…Hypothetically, some variants may be expected to possess differences in various aspects of their poly-modal regulation and exogenous agents, such as zinc ions (Zn 2 + ), may modify their behaviour [10], perhaps with relevance to particular physiological circumstances [11]. However, the functional properties of most of the variants have not been reported.…”

Section: Introductionmentioning

confidence: 99%

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Genetic variants affecting human TRPA1 or TRPM8 structure can be classified in vitro as ‘well expressed’, ‘poorly expressed’ or ‘salvageable’

2015

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SynopsisMultiple mis-sense variants of TRPA1 (transient receptor potential A1) and TRPM8 (transient receptor potential M8) are recorded in the human genome single nt polymorphism (SNP) database, but their potential impact on channel signalling in patho-physiology is not fully explored. Variants, mostly quite rare in the general human population, alter sites in different structural domains of these homo-tetrameric ion channel proteins. The effects of individual SNPs affecting the large cytoplasmic N-terminal domain have not been completely documented for TRPM8 or TRPA1. We examined the Ca 2 + signalling properties of a short-list of eight variants affecting the N-terminal domain by individual expression in human embryonic kidney HEK293 or neuroblastoma (SH-SY5Y) cell lines (four SNP variants for TRPM8: G150R, K423N, R475C, R485W and four for TRPA1: Y69C, A366D, E477K, D573A). These were compared with TRPA1 SNP variants affecting intracellular loops located beyond the N-terminal domain and associated with gain of function (such as increased sensitivity to agonists: TRPA1 R797T and N855S). A substitution in TRPA1 (Y69C) exhibited high expression/sensitivity to agonists (high iCa 2 + max (maximum level of intracellular calcium ion), similar to R797T, but less sensitive than N855S), whereas each of the other non-conservative substitutions exhibited poor signalling response (low iCa 2 + max). Responses from these poorly expressed variants could be salvaged, to different extents, by pre-treating cells with the Src (Src protein) family inhibitor protein kinase inhibitor PP2 (PP2: 4-Amino-3-(4-chlorophenyl)-1-(t-butyl)-1H-pyrazolo [3,4-d]pyrimidine, 4-Amino-5-(4-chlorophenyl)-7-(tbutyl)pyrazolo [3,4-d]pyrimidine), or with micromolar Zn 2 + . The TRPA1 variants and several experimental mutants (TRPA1 Y97F, Y226F and YY654-655FF) expressed poorly in SH-SY5Y compared with HEK293 cells. More in-depth studies are needed to identify SNP variants eliciting gain of function in these TRP (transient receptor potential) channels and to assess their roles in medical conditions.

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Section: Discussionmentioning

confidence: 83%

Section: Introductionmentioning

confidence: 99%

Genetic variants affecting human TRPA1 or TRPM8 structure can be classified in vitro as ‘well expressed’, ‘poorly expressed’ or ‘salvageable’

2015

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“…To elevate cytosolic Ca 2þ by an independent means, cells were transfected with the transient receptor potential channel TrpA1. TrpA1 is a cation permeable channel that can be activated by phytochemicals such as mustard oil and cinnemaldehyde (30), and has been shown to transport both Ca 2þ and Zn 2þ (31). Treatment of cells expressing TrpA1 with the active ingredient in mustard oil, allyl-isothiocyanate (AITC), led to Ca 2þ influx across the plasma membrane and concomitant decrease in ½Zn 2þ ER in accord with the rising ½Ca 2þ cyto (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…S8). Because Zn 2þ also acts as an activator for TrpA1 (31), addition of 100 μM extracellular Zn 2þ in the presence of extracellular Ca 2þ to HeLa cells expressing Fig. S8).…”

Section: Resultsmentioning

confidence: 99%

Measuring steady-state and dynamic endoplasmic reticulum and Golgi Zn ²⁺ with genetically encoded sensors

Qin

Dittmer

Park

et al. 2011

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

279

350

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Zn 2+ plays essential roles in biology, and cells have adopted exquisite mechanisms for regulating steady-state Zn 2+ levels. Although much is known about total Zn 2+ in cells, very little is known about its subcellular distribution. Yet defining the location of Zn 2+ and how it changes with signaling events is essential for elucidating how cells regulate this essential ion. Here we create fluorescent sensors genetically targeted to the endoplasmic reticulum (ER) and Golgi to monitor steady-state Zn 2+ levels as well as flux of Zn 2+ into and out of these organelles. These studies reveal that ER and Golgi contain a concentration of free Zn 2+ that is 100 times lower than the cytosol. Both organelles take up Zn 2+ when cytosolic levels are elevated, suggesting that the ER and Golgi can sequester elevated cytosolic Zn 2+ and thus have the potential to play a role in influencing Zn 2+ toxicity. ER Zn 2+ homeostasis is perturbed by small molecule antagonists of Ca 2+ homeostasis and ER Zn 2+ is released upon elevation of cytosolic Ca 2+ pointing to potential exchange of these two ions across the ER. This study provides direct evidence that Ca 2+ signaling can influence Zn 2+ homeostasis and vice versa, that Zn 2+ dynamics may modulate Ca 2+ signaling.

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“…Three cysteine residues in mouse TRPA1 (Cys415, Cys422, and Cys622, conserved in the human homolog as Cys414, Cys421, and Cys621) were independently identifi ed as the target sites for AITC and cinnamaldehyde [ 31 ]. Intracellular Zn 2+ also activates human TRPA1 by interacting with Cys641 and C-terminal Cys1021/His983 [ 78 ]. Systematic evaluation of TRP channels was performed using a series of reactive disulfi des, such as bis(5-nitro-2-pyridyl) disulfi de and diallyl disulfi de [ 33 ].…”

Section: Modulation Of Trpa1 Channel Activitymentioning

confidence: 99%

TRP Channels: Their Function and Potentiality as Drug Targets

Nishida

Kuwahara

Kozai

et al. 2015

Innovative Medicine

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The transient receptor potential (TRP) proteins are a family of ion channels that act as cellular sensors as well as signal integrators. Several members of the TRP family are sensitive to changes in cellular redox status. Among them, TRPA1 is remarkably susceptible to various oxidants and is known to mediate neuropathic pain and respiratory, vascular, and gastrointestinal functions, making TRPA1 an attractive therapeutic target. However, a method to achieve selective modulation of TRPA1 by small molecules has not yet been established. Most recently, we found that a novel N -nitrosamine compound activates TRPA1 by S -nitrosylation (the addition of a nitric oxide (NO) group to cysteine thiol) and does so with signifi cant selectivity over other NO-sensitive TRP channels. It is proposed that this subtype selectivity is conferred through synergistic effects of electrophilic cysteine transnitrosylation and molecular recognition of the non-electrophilic moiety on the N -nitrosamine. On the other hand, TRPCs are typical receptor-activated Ca 2+ -permeable cation channels, which sense messenger molecules generated downstream of phospholipase activation. Previously, activation of TRPC3 and TRPC6 by diacylglycerol has been reported to play important roles in the pathogenesis of cardiac hypertrophy. Also, a pyrazole compound, Pyr3, which selectively inhibits TRPC3, suppresses cardiac hypertrophy in animal models in vitro and in vivo. We have most recently found that Pyr3 and related compounds are effective in suppressing cardiac fi brosis and ischemia responsible for cardiac remodeling as well. Thus, in this chapter, we describe the molecular pharmacology of TRP modulators and discuss their modulatory mechanisms and pharmacological actions.

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Zinc activates damage-sensing TRPA1 ion channels

Cited by 200 publications

References 49 publications

Genetic variants affecting human TRPA1 or TRPM8 structure can be classified in vitro as ‘well expressed’, ‘poorly expressed’ or ‘salvageable’

Genetic variants affecting human TRPA1 or TRPM8 structure can be classified in vitro as ‘well expressed’, ‘poorly expressed’ or ‘salvageable’

Measuring steady-state and dynamic endoplasmic reticulum and Golgi Zn ²⁺ with genetically encoded sensors

TRP Channels: Their Function and Potentiality as Drug Targets

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Zinc activates damage-sensing TRPA1 ion channels

Cited by 200 publications

References 49 publications

Genetic variants affecting human TRPA1 or TRPM8 structure can be classified in vitro as ‘well expressed’, ‘poorly expressed’ or ‘salvageable’

Genetic variants affecting human TRPA1 or TRPM8 structure can be classified in vitro as ‘well expressed’, ‘poorly expressed’ or ‘salvageable’

Measuring steady-state and dynamic endoplasmic reticulum and Golgi Zn 2+ with genetically encoded sensors

TRP Channels: Their Function and Potentiality as Drug Targets

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Measuring steady-state and dynamic endoplasmic reticulum and Golgi Zn ²⁺ with genetically encoded sensors