Vanilloid-dependent TRPV1 opening trajectory from cryoEM ensemble analysis

Kwon, Do Hoon; Zhang, Feng; Fedor, Justin G.; Suo, Yang; Lee, Seok-Yong

doi:10.1038/s41467-022-30602-2

Cited by 38 publications

(39 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While TRPV1 activation by RTX exhibited similar progressive shifting towards the open state, dwell-times in the closed and open states appeared to be long enough to skip the filtering effect on current amplitude. The allosteric behavior described in the present study may help correlate published high-resolution channel structures (15,32) with distinct functional states towards a better understanding of the activation mechanism of TRPV1 and other ion channels.…”

Section: Resultsmentioning

confidence: 75%

“…Concerted pore opening has been previously proposed for other channel types including Kv channels, for which independent voltage-sensor movements in the four subunits precede channel activation (28,29). Very brief (microseconds) subconductance states were found to be associated with asymmetrical subunit activations by voltage, hence representing intermediate gating states (30,31) (15,32) with distinct functional states towards a better understanding of the activation mechanism of TRPV1 and other ion channels.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

TRPV1 Opening is Stabilized Equally by Its Four Subunits

Nguyen

et al. 2023

Preprint

View full text Add to dashboard Cite

Capsaicin receptor TRPV1 is a nociceptor for vanilloid molecules such as capsaicin and resiniferatoxin (RTX). Even though cryo-EM structures of TRPV1 in complex with these molecules are available, how their binding energetically favors the open conformation is not known. Here we report an approach to control the number of bound RTX molecules (0-to-4) in functional mouse TRPV1. The approach allowed direct measurements of each of the intermediate open states under equilibrium conditions at both macroscopic and single-molecule levels. We found that RTX binding to each of the four subunits contributes virtually the same activation energy, which we estimated to be 1.86 kcal/mol and found to arise predominately from destabilizing the closed conformation. We further showed that sequential bindings of RTX increase open probability without altering single-channel conductance, confirming that there is likely a single open-pore conformation for TRPV1 activated by RTX.

show abstract

Section: Resultsmentioning

confidence: 75%

Section: Resultsmentioning

confidence: 99%

TRPV1 Opening is Stabilized Equally by Its Four Subunits

Nguyen

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…This suggests that balancing the two different neurophysiological signaling pathways (calcium signaling and actin cytoskeleton remodeling) requires fine tuning of the TRPV4-RhoA interaction. TRPV channels are highly allosterically coupled across their domains, as was previously shown 40 , and because we observed progressive changes in RhoA occupancy during gating we attempted to infer the effect of RhoA on TRPV4 gating. Structural alignment at the TMDs show that each ankyrin repeat (AR) in the ARD swings toward the membrane by ~6 Å from the closed to the open state (Fig.…”

Section: Neuropathy Mutations Disrupt Rhoa Binding To Trpv4mentioning

confidence: 85%

Structural insights into TRPV4-Rho GTPase signaling complex function and disease

Kwon

Zhang

McCray

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Crosstalk between ion channels and small GTPases is critical during homeostasis and disease, but little is known about the structural underpinnings of these interactions. TRPV4 is a polymodal, calcium-permeable cation channel that has emerged as a potential therapeutic target in multiple conditions. Gain-of-function mutations also cause hereditary neuromuscular disease. Here, we present cryo-EM structures of human TRPV4 in complex with RhoA in the apo, antagonist-bound closed, and agonist-bound open states. These structures reveal the mechanism of ligand-dependent TRPV4 gating. Channel activation is associated with rigid-body rotation of the intracellular ankyrin repeat domain, but state-dependent interaction with membrane-anchored RhoA constrains this movement. Notably, many residues at the TRPV4-RhoA interface are mutated in disease and perturbing this interface by introducing mutations into either TRPV4 or RhoA increases TRPV4 channel activity. Together, these results suggest that the interaction strength between TRPV4 and RhoA tunes TRPV4-mediated calcium homeostasis and actin remodeling, and that disruption of TRPV4-RhoA interactions leads to TRPV4-related neuromuscular disease, findings that will guide TRPV4 therapeutics development.

show abstract

“…This promotes selectivity filter dilation and pore loop rearrangements. 42,80 The tail is a very flexible fatty acyl chain in capsaicin, which adopt more than one fixed conformation and contributed to the lack of clear electron density cryo-EM data in previous studies. 81,82 Thermo-TRPV channels are essentially non-selective cation channels allowing the permeation of bulky organic cations through conformational rearrangements, this in part at the level of the outer pore domain, as a response to the interaction with RTx.…”

Section: Discussionmentioning

confidence: 99%

Non-canonical helical transitions and conformational switching are associated with characteristic flexibility profiles in ligand-gated TRP channels and voltage-dependent Kv channels

Balleza

García-Morales

2022

Preprint

View full text Add to dashboard Cite

The transient receptor potential (TRP) superfamily of cation channels is divided into several subfamilies. The TRPV (vanilloid) subfamily is composed of proteins that undergo a closed-to-open gating transition in response to various physical and chemical stimuli, including heat transfer phenomena. TRPV1, the prototypical member of these thermo-TRP channels, is activated under physiological conditions by noxious high temperatures (>42°C), and by pungent chemicals including capsaicin and resiniferatoxin . Like TRPV1, TRPV2 is also activated at >52°C as well as 2-aminoethoxydiphenyl borate and cannabidiol. On the other hand, the main stimulus for Kv channels is the potential difference across the membrane. Notwithstanding these differences in their activation temperatures, ligand- or voltage-dependence, the gating mechanisms of these channels exhibit conformational transition pathways in common. Understanding these conformational changes in terms of the sequence determinants underlying these structural transitions helps to reveal residues with great functional relevance. This work examines the side-chain flexibility in regions undergoing peculiar helical transitions. We found that α-to-π helical transitions are associated with patterns of local rigidity whereas α-to-3 transitions are mainly associated with high local flexibility profiles. We also study the relationship between flexibility and protein order, both in these dynamic regions and in the rest of the transmembrane domains of these proteins. Our analysis shows that flexibility and protein disorder are two complementary parameters that could reveal conformational heterogeneity and the dynamic behavior of specific segments.

show abstract

Vanilloid-dependent TRPV1 opening trajectory from cryoEM ensemble analysis

Cited by 38 publications

References 52 publications

TRPV1 Opening is Stabilized Equally by Its Four Subunits

TRPV1 Opening is Stabilized Equally by Its Four Subunits

Structural insights into TRPV4-Rho GTPase signaling complex function and disease

Non-canonical helical transitions and conformational switching are associated with characteristic flexibility profiles in ligand-gated TRP channels and voltage-dependent Kv channels

Contact Info

Product

Resources

About