Identification of Molecular Determinants for a Potent Mammalian TRPA1 Antagonist by Utilizing Species Differences

Nakatsuka, Kazumasa; Gupta, Rupai; Saito, Shigeru; Banzawa, Nagako; Takahashi, Kenji; Tominaga, Makoto; Ohta, Toshio

doi:10.1007/s12031-013-0060-2

Cited by 32 publications

(27 citation statements)

References 31 publications

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“…It has been shown that two amino acid residues located in the predicted inner site of TM5 are critical for the antagonistic action of AP18 (26). Similarly, we recently reported that these two amino acid residues (serine 873 and tyrosine 874) on human TRPA1 were also important for the antagonistic effect of A967079 (29). Furthermore, by analyses using a homology model of TRPA1 based on Kv1.2, it has also been reported that phenylalanine 944, valine 948, and isoleucine 950 in the predicted loop between TM5 and TM6 are involved in the antagonistic action of A967079 (38).…”

Section: Discussionsupporting

confidence: 53%

“…By utilizing species differences, we identified two amino acid residues located within the putative fifth transmembrane (TM5) domain as critical determinants for the antagonistic action of A967079 (29). Quite recently, we also analyzed functional properties of chicken TRPA1, and we reported that it is a heat sensor, but not a cold one, unlike rodent TRPA1 (30).…”

Section: Trpa1mentioning

confidence: 99%

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Molecular Basis Determining Inhibition/Activation of Nociceptive Receptor TRPA1 Protein

Banzawa

Saito

Imagawa

et al. 2014

Journal of Biological Chemistry

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Background: Nociceptive TRPA1 is an important drug discovery target for identification of analgesics. Results: A967079, one of the most potent mammalian TRPA1 antagonists, showed marked species differences. Conclusion: A single amino acid was responsible for inhibitory/stimulatory actions of A967079. Significance: The present data provide novel insight in the search for analgesics targeting TRPA1.

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Section: Discussionsupporting

confidence: 53%

Section: Trpa1mentioning

confidence: 99%

Molecular Basis Determining Inhibition/Activation of Nociceptive Receptor TRPA1 Protein

Banzawa

Saito

Imagawa

et al. 2014

Journal of Biological Chemistry

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“…S2d,e). CA concentrations higher than the reported EC 50 values (0.13 mM and 0.39 mM for hTRPA1 and fTRPA1, respectively) were chosen1013, and we waited for the CA-evoked currents to be stabilized or desensitized. Figure 1a and b shows the reversible current responses of wild type hTRPA1 (wt-hTRPA1) and fTRPA1 (wt-fTRPA1) to repeated application of 0.3 and 0.5 mM CA, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…For example, A967079 (A96) and AP18, both of which are structurally similar, are potent mammalian TRPA1 antagonists, although their inhibitory effects on TRPA1 vary among species101112. Comparative and mutagenesis experiments with TRPA1 from different species revealed that several amino acids in the TM5 domain are crucial to the effects of these two antagonists91213. Furthermore, a recent study reporting the detailed structure of human TRPA1 identified a binding site for A96 that is in the vicinity of sites found previously14.…”

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confidence: 99%

Structural basis of TRPA1 inhibition by HC-030031 utilizing species-specific differences

Gupta

Saito

Mori

et al. 2016

Sci Rep

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Pain is a harmful sensation that arises from noxious stimuli. Transient receptor potential ankyrin 1 (TRPA1) is one target for studying pain mechanisms. TRPA1 is activated by various stimuli such as noxious cold, pungent natural products and environmental irritants. Since TRPA1 is an attractive target for pain therapy, a few TRPA1 antagonists have been developed and some function as analgesic agents. The responses of TRPA1 to agonists and antagonists vary among species and these species differences have been utilized to identify the structural basis of activation and inhibition mechanisms. The TRPA1 antagonist HC-030031 (HC) failed to inhibit frog TRPA1 (fTRPA1) and zebrafish TRPA1 activity induced by cinnamaldehyde (CA), but did inhibit human TRPA1 (hTRPA1) in a heterologous expression system. Chimeric studies between fTRPA1 and hTRPA1, as well as analyses using point mutants, revealed that a single amino acid residue (N855 in hTRPA1) significantly contributes to the inhibitory action of HC. Moreover, the N855 residue and the C-terminus region exhibited synergistic effects on the inhibition by HC. Molecular dynamics simulation suggested that HC stably binds to hTRPA1-N855. These findings provide novel insights into the structure-function relationship of TRPA1 and could lead to the development of more effective analgesics targeted to TRPA1.

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“…Electrophilic compounds are known to activate TRPA1 through covalent modification of cysteines and a lysine in the N-terminal region (12,13). In addition, the S5/S6 domain may serve as a target region for nonelectrophilic agonists, including menthol (14), eudesmol (15), and protons (16), or several antagonists (17,18), including the potent inhibitor A-967079 (14,(19)(20)(21). Furthermore, a recent electron cryo-microscopy (cryo-EM) study (22) has revealed the high-resolution structure of TRPA1 bound to an agonist or antagonists.…”

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confidence: 99%

Identification of a putative binding site critical for general anesthetic activation of TRPA1

Ton

Phan

Abramyan

et al. 2017

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

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General anesthetics suppress CNS activity by modulating the function of membrane ion channels, in particular, by enhancing activity of GABA A receptors. In contrast, several volatile (isoflurane, desflurane) and i.v. (propofol) general anesthetics excite peripheral sensory nerves to cause pain and irritation upon administration. These noxious anesthetics activate transient receptor potential ankyrin repeat 1 (TRPA1), a major nociceptive ion channel, but the underlying mechanisms and site of action are unknown. Here we exploit the observation that pungent anesthetics activate mammalian but not Drosophila TRPA1. Analysis of chimeric Drosophila and mouse TRPA1 channels reveal a critical role for the fifth transmembrane domain (S5) in sensing anesthetics. Interestingly, we show that anesthetics share with the antagonist A-967079 a potential binding pocket lined by residues in the S5, S6, and the first pore helix; isoflurane competitively disrupts A-967079 antagonism, and introducing these mammalian TRPA1 residues into dTRPA1 recapitulates anesthetic agonism. Furthermore, molecular modeling predicts that isoflurane and propofol bind to this pocket by forming H-bond and halogenbond interactions with Ser-876, Met-915, and Met-956. Mutagenizing Met-915 or Met-956 selectively abolishes activation by isoflurane and propofol without affecting actions of A-967079 or the agonist, menthol. Thus, our combined experimental and computational results reveal the potential binding mode of noxious general anesthetics at TRPA1. These data may provide a structural basis for designing drugs to counter the noxious and vasorelaxant properties of general anesthetics and may prove useful in understanding effects of anesthetics on related ion channels.

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Identification of Molecular Determinants for a Potent Mammalian TRPA1 Antagonist by Utilizing Species Differences

Cited by 32 publications

References 31 publications

Molecular Basis Determining Inhibition/Activation of Nociceptive Receptor TRPA1 Protein

Molecular Basis Determining Inhibition/Activation of Nociceptive Receptor TRPA1 Protein

Structural basis of TRPA1 inhibition by HC-030031 utilizing species-specific differences

Identification of a putative binding site critical for general anesthetic activation of TRPA1

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