Mutagenesis and Temperature-Sensitive Little Machines

Pertusa, Marı́a; Moldenhauer, Hans; Brauchi, Sebastián; Latorre, Ramón; Madrid, Rodolfo; Orio, Patricio

doi:10.5772/50333

Cited by 5 publications

(8 citation statements)

References 107 publications

(164 reference statements)

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“…Cooperativity intrinsic to TRP channels may result in allosteric coupling of distinct activation stimuli. A good example of the allosteric nature of TRP channels would be the case of temperature-activated TRP channels (thermoTRPs) (Latorre et al, 2007 ; Pertusa et al, 2012 ).…”

Section: Trp Channelsmentioning

confidence: 99%

A structural view of ligand-dependent activation in thermoTRP channels

2014

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Transient Receptor Potential (TRP) proteins are a large family of ion channels, grouped into seven sub-families. Although great advances have been made regarding the activation and modulation of TRP channel activity, detailed molecular mechanisms governing TRP channel gating are still needed. Sensitive to electric, chemical, mechanical, and thermal cues, TRP channels are tightly associated with the detection and integration of sensory input, emerging as a model to study the polymodal activation of ion channel proteins. Among TRP channels, the temperature-activated kind constitute a subgroup by itself, formed by Vanilloid receptors 1–4, Melastatin receptors 2, 4, 5, and 8, TRPC5, and TRPA1. Some of the so-called “thermoTRP” channels participate in the detection of noxious stimuli making them an interesting pharmacological target for the treatment of pain. However, the poor specificity of the compounds available in the market represents an important obstacle to overcome. Understanding the molecular mechanics underlying ligand-dependent modulation of TRP channels may help with the rational design of novel synthetic analgesics. The present review focuses on the structural basis of ligand-dependent activation of TRPV1 and TRPM8 channels. Special attention is drawn to the dissection of ligand-binding sites within TRPV1, PIP2-dependent modulation of TRP channels, and the structure of natural and synthetic ligands.

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Section: Trp Channelsmentioning

confidence: 99%

A structural view of ligand-dependent activation in thermoTRP channels

2014

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“…Whole-cell recordings were performed at 14, 22, and 28°C ( Figure 5E), producing an evident increase in the macroscopic current ( Figures 5E and 5F). It is important to note that although the calculated temperature dependence of the channel allows it to be classified as temperature-dependent (Q 10 = 5, n = 5; Figure 5G), the activation coefficient was modest when compared with the mammalian thermoTRP set (Q 10 > 10; Pertusa et al, 2012).…”

Section: Cloning Heterologous Expression and Biophysical Characterimentioning

confidence: 99%

“…A subset of TRP channels, dubbed thermoTRPs, display an exquisite thermal sensitivity, often having Q 10 values over 10 (Clapham, 2009;Latorre et al, 2009;Pertusa et al, 2012). As several channels from the C, M, and V families have been described as temperature-activated channels (Pertusa et al, 2012), we analyzed Cr-TRP1's temperature sensitivity.…”

Section: Cloning Heterologous Expression and Biophysical Characterimentioning

confidence: 99%

“…As several channels from the C, M, and V families have been described as temperature-activated channels (Pertusa et al, 2012), we analyzed Cr-TRP1's temperature sensitivity. Whole-cell recordings were performed at 14, 22, and 28°C ( Figure 5E), producing an evident increase in the macroscopic current ( Figures 5E and 5F).…”

Section: Cloning Heterologous Expression and Biophysical Characterimentioning

confidence: 99%

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A Transient Receptor Potential Ion Channel in Chlamydomonas Shares Key Features with Sensory Transduction-Associated TRP Channels in Mammals

et al. 2015

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Sensory modalities are essential for navigating through an ever-changing environment. From insects to mammals, transient receptor potential (TRP) channels are known mediators for cellular sensing. Chlamydomonas reinhardtii is a motile single-celled freshwater green alga that is guided by photosensory, mechanosensory, and chemosensory cues. In this type of alga, sensory input is first detected by membrane receptors located in the cell body and then transduced to the beating cilia by membrane depolarization. Although TRP channels seem to be absent in plants, C. reinhardtii possesses genomic sequences encoding TRP proteins. Here, we describe the cloning and characterization of a C. reinhardtii version of a TRP channel sharing key features present in mammalian TRP channels associated with sensory transduction. In silico sequence-structure analysis unveiled the modular design of TRP channels, and electrophysiological experiments conducted on Human Embryonic Kidney-293T cells expressing the Cr-TRP1 clone showed that many of the core functional features of metazoan TRP channels are present in Cr-TRP1, suggesting that basic TRP channel gating characteristics evolved early in the history of eukaryotes.

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“…This protein belongs to the family of transient receptor potential (TRP) 2 ion channels, of which another 10 thermosensitive channels have been identified: TRPV1-4, TRPA1, TRPM2-5 and TRPC5 (reviewed in Refs. [7][8][9]. TRPM8 is activated by cold and by cooling agents such as menthol and icilin (10,11), it displays weak voltage dependence (12)(13)(14), and it has a key role not only in cold sensing but also in cancer and pathological cold-induced pain (reviewed in Refs.…”

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confidence: 99%

Critical role of the pore domain in the cold response of TRPM8 channels identified by ortholog functional comparison

Pertusa

Rivera

González

et al. 2018

Journal of Biological Chemistry

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In mammals, the main molecular entity involved in innocuous cold transduction is TRPM8. This polymodal ion channel is activated by cold, cooling compounds such as menthol and voltage. Despite its relevance, the molecular determinants involved in its activation by cold remain elusive. In this study we explored the use of TRPM8 orthologs with different cold responses as a strategy to identify new molecular determinants related with their thermosensitivity. We focused on mouse TRPM8 (mTRPM8) and chicken TRPM8 (cTRPM8), which present complementary thermosensitive and chemosensitive phenotypes. Although mTRPM8 displays larger responses to cold than cTRPM8 does, the avian ortholog shows a higher sensitivity to menthol compared with the mouse channel, in both HEK293 cells and primary somatosensory neurons. We took advantage of these differences to build multiple functional chimeras between these orthologs, to identify the regions that account for these discrepancies. Using a combination of calcium imaging and patch clamping, we identified a region encompassing positions 526-556 in the N terminus, whose replacement by the cTRPM8 homolog sequence potentiated its response to agonists. More importantly, we found that the characteristic cold response of these orthologs is due to nonconserved residues located within the pore loop, suggesting that TRPM8 has evolved by increasing the magnitude of its cold response through changes in this region. Our results reveal that these structural domains are critically involved in cold sensitivity and functional modulation of TRPM8, and support the idea that the pore domain is a key molecular determinant in temperature responses of this thermo-transient receptor potential (TRP) channel.

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Mutagenesis and Temperature-Sensitive Little Machines

Cited by 5 publications

References 107 publications

A structural view of ligand-dependent activation in thermoTRP channels

A structural view of ligand-dependent activation in thermoTRP channels

A Transient Receptor Potential Ion Channel in Chlamydomonas Shares Key Features with Sensory Transduction-Associated TRP Channels in Mammals

Critical role of the pore domain in the cold response of TRPM8 channels identified by ortholog functional comparison

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