The N-terminal Ankyrin Repeat Domain Is Not Required for Electrophile and Heat Activation of the Purified Mosquito TRPA1 Receptor

Survery, Sabeen; Moparthi, Lavanya; Kjellbom, Per; Högestätt, Edward D.; Zygmunt, Peter M.; Johanson, Urban

doi:10.1074/jbc.m116.743443

Cited by 18 publications

(35 citation statements)

References 51 publications

(109 reference statements)

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“…Previous studies have shown that species differences between a small number of residues in the transmembrane core give rise to capsaicin sensitivity in TRPV1 and icilin sensitivity in TRPM8 (19,25). Additionally, the transmembrane domain of TRPA1 is sufficient to recapitulate temperature and agonist response of this channel (46,47). Given the importance of the transmembrane core in TRP channel function, we focused our search on this region.…”

Section: Species-dependent Modulation Of Trpm8 By Pirtmentioning

confidence: 99%

Phosphoinositide-interacting regulator of TRP (PIRT) has opposing effects on human and mouse TRPM8 ion channels

Hilton

Salehpour

Sisco

et al. 2018

Journal of Biological Chemistry

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Transient receptor potential melastatin 8 (TRPM8) is a cold-sensitive ion channel with diverse physiological roles. TRPM8 activity is modulated by many mechanisms, including an interaction with the small membrane protein phosphoinositide-interacting regulator of TRP (PIRT). Here, using comparative electrophysiology experiments, we identified species-dependent differences between the human and mouse TRPM8-PIRT complexes. We found that human PIRT attenuated human TPRM8 conductance, unlike mouse PIRT, which enhanced mouse TRPM8 conductance. Quantitative Western blot analysis demonstrates that this effect does not arise from decreased trafficking of TRPM8 to the plasma membrane. Chimeric human/mouse TRPM8 channels were generated to probe the molecular basis of the PIRT modulation, and the effect was recapitulated in a pore domain chimera, demonstrating the importance of this region for PIRT-mediated regulation of TRPM8. Moreover, recombinantly expressed and purified human TRPM8 S1-S4 domain (comprising transmembrane helices S1-S4, also known as the sensing domain, ligand-sensing domain, or voltage sensing-like domain) and full-length human PIRT were used to investigate binding between the proteins. NMR experiments, supported by a pulldown assay, indicated that PIRT binds directly and specifically to the TRPM8 S1-S4 domain. Binding became saturated as the S1-S4:PIRT mole ratio approached 1. Our results have uncovered species-specific TRPM8 modulation by PIRT. They provide evidence for a direct interaction between PIRT and the TRPM8 S1-S4 domain with a 1:1 binding stoichiometry, suggesting that a functional tetrameric TRPM8 channel has four PIRT-binding sites.

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Section: Species-dependent Modulation Of Trpm8 By Pirtmentioning

confidence: 99%

Phosphoinositide-interacting regulator of TRP (PIRT) has opposing effects on human and mouse TRPM8 ion channels

Hilton

Salehpour

Sisco

et al. 2018

Journal of Biological Chemistry

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“…Presumptive IR coreceptors (IRco-y) are required for the reception of multiple chemical classes in multiple cell types ( Abuin et al, 2011 ; Pitts et al, 2017 ), but their exact role in signal transduction remains elusive. Transient Receptor Potential (TRP) channels assemble as homotetramers ( Survery et al, 2016 ) sensitive to electrophiles (allyl isothiocyanate shown as example). These multimodal channels are approximately three times as large as ORs and GRs.…”

Section: Primary Receptor Familiesmentioning

confidence: 99%

“…Together, these findings support the notion that Ag TRPA1 functions as a peripheral thermoreceptor in mosquito antenna. More recent work has uncovered an additional chemosensory role for Ag TRPA1 ( Survery et al, 2016 ). Patch clamp recordings on heterologously expressed and purified, full-length Ag TRPA1 and truncated Δ1–776 Ag TRPA1 (lacking the N-terminal ARD) demonstrated that both proteins are functional, as each responded to the electrophilic compounds, allyl isothiocyanate and cinnamaldehyde, as well as heat.…”

Section: Ancillary Membrane Proteinsmentioning

confidence: 99%

“…Patch clamp recordings on heterologously expressed and purified, full-length Ag TRPA1 and truncated Δ1–776 Ag TRPA1 (lacking the N-terminal ARD) demonstrated that both proteins are functional, as each responded to the electrophilic compounds, allyl isothiocyanate and cinnamaldehyde, as well as heat. Their similar intrinsic fluorescence properties and related quenching of tryptophan, when activated by allyl isothiocyanate or heat, led the researchers to conclude that conformational change in the lipid bilayer occurs independently and outside of the N-terminal domain ( Survery et al, 2016 ). As such, Ag TRPA1 is both a thermo- and chemoreceptor, and while the N-terminal domain’s function is unknown, it is hypothesized to play a role in tuning the channel’s response ( Survery et al, 2016 ).…”

Section: Ancillary Membrane Proteinsmentioning

confidence: 99%

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Membrane Proteins Mediating Reception and Transduction in Chemosensory Neurons in Mosquitoes

et al. 2018

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Mosquitoes use chemical cues to modulate important behaviors such as feeding, mating, and egg laying. The primary chemosensory organs comprising the paired antennae, maxillary palps and labial palps are adorned with porous sensilla that house primary sensory neurons. Dendrites of these neurons provide an interface between the chemical environment and higher order neuronal processing. Diverse proteins located on outer membranes interact with chemicals, ions, and soluble proteins outside the cell and within the lumen of sensilla. Here, we review the repertoire of chemosensory receptors and other membrane proteins involved in transduction and discuss the outlook for their functional characterization. We also provide a brief overview of select ion channels, their role in mammalian taste, and potential involvement in mosquito taste. These chemosensory proteins represent targets for the disruption of harmful biting behavior and disease transmission by mosquito vectors.

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“…Thus, selective blockers are added to confirm that currents are due to activity of a reconstituted channel. It is reported that reconstituted TRPA1 is blocked by non‐selective TRP channel pore blocker ruthenium red (Moparthi et al., ; Survery et al., ) as well as by the selective TRPA1 antagonist HC030031 (Moparthi et al., ) in the Port‐a‐Patch system.…”

Section: Commentarymentioning

confidence: 99%

Reconstitution and Electrophysiological Characterization of Ion Channels in Lipid Bilayers

et al. 2018

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Detergent-solubilized purified ion channels can be reconstituted into lipid bilayers for electrophysiological analysis. Traditionally, ion channels were inserted into vesicles and subsequently fused with planar "black lipid membranes" formed from lipids dissolved in a hydrophobic solvent such as decane. Provided in this article is a step-by-step guide to reconstitute purified ion channel proteins into giant unilamellar vesicles (GUVs). This procedure results in the formation of proteoliposomes that can be used for planar bilayer formation and electrophysiological characterization of single-channel currents. By using preformed GUVs it is possible to omit the membrane solvent. Compared to traditional preparations, the lipid bilayers formed from GUVs provide an environment that more closely resembles the native cell membrane. Also described is an alternate protocol that entails the production of planar lipid bilayers from GUVs onto which proteins in detergent are added. © 2018 by John Wiley & Sons, Inc.

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The N-terminal Ankyrin Repeat Domain Is Not Required for Electrophile and Heat Activation of the Purified Mosquito TRPA1 Receptor

Cited by 18 publications

References 51 publications

Phosphoinositide-interacting regulator of TRP (PIRT) has opposing effects on human and mouse TRPM8 ion channels

Phosphoinositide-interacting regulator of TRP (PIRT) has opposing effects on human and mouse TRPM8 ion channels

Membrane Proteins Mediating Reception and Transduction in Chemosensory Neurons in Mosquitoes

Reconstitution and Electrophysiological Characterization of Ion Channels in Lipid Bilayers

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