Exploring the activation mechanism of TRPM8 channel by targeted MD simulations

Pedretti, Alessandro; Labozzetta, Alessandra; Monte, Matteo Lo; Beccari, Andrea R.; Moriconi, Alessio; Vistoli, Giulio

doi:10.1016/j.bbrc.2011.08.134

Cited by 14 publications

(9 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These data demonstrate that the N-x-x-D motif in fact is essential for the sensitivity of TRPM8 to icilin but is also critical for the responses to menthol and to cold, and therefore experimentally support the recently proposed model [11] based on molecular dynamic simulations, strongly suggesting that S3 is essential for the activation of TRPM8 by menthol.…”

Section: Resultssupporting

confidence: 86%

“…Since electrostatic interactions of this motif with other transmembrane segments have been proposed [11], we swapped the position of the outer residues of the N-x-x-D motif or altered the hydrophobicity of the inner residues. We report strikingly differential effects on the responses to menthol and cold of TRPM8 and to ADP-ribose (ADPR) of TRPM2, reflecting the different modes of activation in spite of common essential structural elements.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Importance of a Conserved Sequence Motif in Transmembrane Segment S3 for the Gating of Human TRPM8 and TRPM2

et al. 2012

View full text Add to dashboard Cite

For mammalian TRPM8, the amino acid residues asparagine-799 and aspartate-802 are essential for the stimulation of the channel by the synthetic agonist icilin. Both residues belong to the short sequence motif N-x-x-D within the transmembrane segment S3 highly conserved in the entire superfamily of voltage-dependent cation channels, among them TRPM8. Moreover, they are also conserved in the closely related TRPM2 channel, which is essentially voltage-independent. To analyze the differential roles of the motif for the voltage-dependent and voltage-independent gating, we performed reciprocal replacements of the asparagine and aspartate within the S3 motif in both channels, following the proposed idea that specific electrostatic interactions with other domains take place during gating. Wild-type and mutant channels were heterologeously expressed in HEK-293 cells and channel function was analyzed by whole-cell patch-clamp analysis as well as by Ca2+-imaging. Additionally, the expression of the channels in the plasma membrane was tested by Western blot analysis, in part after biotinylation. For the mutations of TRPM8, responses to menthol were only compromised if also the expression of the glycosylated channel isoform was prevented. In contrast, responses to cold were consistently and significantly attenuated but not completely abolished. For TRPM2, surface expression was not significantly affected by any of the mutations but channel function was only retained in one variant. Remarkably, this was the variant of which the corresponding mutation in TRPM8 exerted the most negative effects both on channel function and expression. Furthermore, we performed an exchange of the inner pair of residues of the N-x-x-D motif between the two channels, which proved deleterious for the functional expression of TRPM8 but ineffective on TRPM2. In conclusion, the N-x-x-D motif plays specific roles in TRPM8 and TRPM2, reflecting different requirements for voltage-dependent and voltage-independent channel gating.

show abstract

Section: Resultssupporting

confidence: 86%

Section: Introductionmentioning

confidence: 99%

Importance of a Conserved Sequence Motif in Transmembrane Segment S3 for the Gating of Human TRPM8 and TRPM2

et al. 2012

View full text Add to dashboard Cite

show abstract

“…Molecular dynamics (MD) simulation is the method of choice for exploring protein dynamics and energetics under physiological conditions with atomic resolution ( Karplus and McCammon, 2002 ). MD was used previously to study various ion channels ( Lindahl and Sansom, 2008 ; Nury et al, 2010 , 2011 ; Zhu and Hummer, 2010 ; Dong and Zhou, 2011 ; Delemotte et al, 2012 ; Du et al, 2012 ; Vargas et al, 2012 ; Calimet et al, 2013 ), including truncated domains ( Sotomayor et al, 2005 ; Susankova et al, 2007 ) and homology models ( Brauchi et al, 2007 ; Susankova et al, 2007 ; Pedretti et al, 2011 ) of TRP channels. However, MD simulation is computationally expensive.…”

Section: Introductionmentioning

confidence: 99%

A combined coarse-grained and all-atom simulation of TRPV1 channel gating and heat activation

Zheng

Feng

2015

Journal of General Physiology

View full text Add to dashboard Cite

show abstract

“…Structurally, each of the four TRPM8 subunits has six transmembranous helixes, S1–6, where S1–4 comprise the voltage‐sensitive unit. TRPM8 agonists bind to S2 causing S4 elongation and eventually a conformational shift leading to channel opening (Pedretti et al., , ). The role of TRPA1 as a noxious cold receptor, co‐localized with TRPV1, was explored 1 year later (Story et al., ).…”

Section: Introductionmentioning

confidence: 99%

A review of topical high‐concentration L‐menthol as a translational model of cold allodynia and hyperalgesia

Andersen

Olsen

Møller

et al. 2013

European Journal of Pain

View full text Add to dashboard Cite

Background: Cold allodynia and cold hyperalgesia are both elusive features of neuropathic pain, particularly in patients with various polyneuropathies. Numerous studies have suggested that topical application of L-menthol causes temporary cold hypersensitivity and thus acts as a proxy for associated symptoms. This review summarizes studies on L-menthol-induced nociception, cold allodynia and cold hyperalgesia in vitro, in animals and in humans. Methods: A comprehensive literature search was performed using the PubMed and Google Scholar databases until February 2013. Obtained manuscripts were reviewed for relevancy and reference lists of the retrieved articles were cross-checked for additional important studies. Solely the literature regarding topical application of L-menthol in humans was attained systematically. Of the total identified studies (96), 10 met the inclusion criteria being controlled studies applying L-menthol at a concentration of ≥30%. Results: The extracted data are meticulously compared and presented with emphasis on clarity and transparency. In seven animal studies, cold allodynia or hyperalgesia was successfully established utilizing various methods. Eight studies in healthy volunteers unanimously reported a significant increase in cold pain threshold, representing cold allodynia and increased supra-threshold cold pain sensitivity, thus demonstrating cold hyperalgesia. Conclusions: Topical high-concentration L-menthol consistently induces cold hypersensitivity in animals and humans, thus constituting a predictable surrogate model of cold allodynia and hyperalgesia. Understanding translational features of this model and its underlying mechanisms could be valuable in preclinical and human phases of drug development and in improving current treatment of patients with polyneuropathy.

show abstract

Exploring the activation mechanism of TRPM8 channel by targeted MD simulations

Cited by 14 publications

References 23 publications

Importance of a Conserved Sequence Motif in Transmembrane Segment S3 for the Gating of Human TRPM8 and TRPM2

Importance of a Conserved Sequence Motif in Transmembrane Segment S3 for the Gating of Human TRPM8 and TRPM2

A combined coarse-grained and all-atom simulation of TRPV1 channel gating and heat activation

A review of topical high‐concentration L‐menthol as a translational model of cold allodynia and hyperalgesia

Contact Info

Product

Resources

About