A single TRPV1 amino acid controls species sensitivity to capsaicin

Chu, Ying; Cohen, Bruce E.; Chuang, Huai-hu

doi:10.1038/s41598-020-64584-2

A single TRPV1 amino acid controls species sensitivity to capsaicin

Journal Article · Fri May 15 00:00:00 EDT 2020 · Scientific Reports

DOI:https://doi.org/10.1038/s41598-020-64584-2· OSTI ID:1631670

Chu, Ying ^[1]; Cohen, Bruce E. ^[2]; Chuang, Huai-hu ^[1]

Academia Sinica, Taipei (Taiwan). Institute of Molecular Biology
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

Chili peppers produce capsaicin (a vanilloid) that activates the transient receptor potential cation channel subfamily V member 1 (TRPV1) on sensory neurons to alter their membrane potential and induce pain. To identify residues responsible for differential TRPV1 capsaicin sensitivity among species, we used intracellular Ca²⁺ imaging to characterize chimeras composed of capsaicin-sensitive rat TRPV1 (rTRPV1) and capsaicin-insensitive chicken TRPV1 (cTRPV1) exposed to a series of capsaicinoids. We found that chimeras containing rat E570-V686 swapped into chicken receptors displayed capsaicin sensitivity, and that simply changing the alanine at position 578 in the S4-S5 helix of the chicken receptor to a glutamic acid was sufficient to endow it with capsaicin sensitivity in the micromolar range. Moreover, introduction of lysine, glutamine or proline at residue A578 also elicited capsaicin sensitivity in cTRPV1. Similarly, replacing corresponding rTRPV1 residue E570 with lysine or glutamine retained capsaicin sensitivity. The hydrophilic capsaicin analog Cap-EA activated a cTRPV1-A578E mutant, suggesting that A578 may participate in vanilloid binding. The hydrophilic vanilloid agonist zingerone did not activate any A578 mutants with capsaicin sensitivity, suggesting that the vanilloid group alone is not sufficient for receptor activation. Our study demonstrates that a subtle modification of TRPV1 in different species globally alters capsaicin responses.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES); Ministry of Science and Technology

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1631670

Journal Information:: Scientific Reports, Journal Name: Scientific Reports Journal Issue: 1 Vol. 10; ISSN 2045-2322

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

References (51)

A new generation of Ca2+ indicators with greatly improved fluorescence properties. Grynkiewicz, G.; Poenie, M.; Tsien, R. Y. Journal of Biological Chemistry, Vol. 260, Issue 6 https://doi.org/10.1016/s0021-9258(19)83641-4	journal	March 1985
Ganglion-specific splicing of TRPV1 underlies infrared sensation in vampire bats Gracheva, Elena O.; Cordero-Morales, Julio F.; González-Carcacía, José A. Nature, Vol. 476, Issue 7358 https://doi.org/10.1038/nature10245	journal	August 2011
Structure of the TRPV1 ion channel determined by electron cryo-microscopy Liao, Maofu; Cao, Erhu; Julius, David Nature, Vol. 504, Issue 7478 https://doi.org/10.1038/nature12822	journal	December 2013
Selective disruption of high sensitivity heat activation but not capsaicin activation of TRPV1 channels by pore turret mutations Cui, Yuanyuan; Yang, Fan; Cao, Xu The Journal of General Physiology, Vol. 139, Issue 4 https://doi.org/10.1085/jgp.201110724	journal	March 2012
Functional Role of C-Terminal Cytoplasmic Tail of Rat Vanilloid Receptor 1 Vlachová, Viktorie; Teisinger, Jan; Sušánková, Klára The Journal of Neuroscience, Vol. 23, Issue 4 https://doi.org/10.1523/jneurosci.23-04-01340.2003	journal	February 2003
Advances in the use of the fluorescent probe fura-2 for the estimation of intrasynatposomal calcium Yates, S. L.; Fluhler, E. N.; Lippiello, P. M. Journal of Neuroscience Research, Vol. 32, Issue 2 https://doi.org/10.1002/jnr.490320215	journal	June 1992
Structural insights into transient receptor potential vanilloid type 1 (TRPV1) from homology modeling, flexible docking, and mutational studies Lee, Jin Hee; Lee, Yoonji; Ryu, HyungChul Journal of Computer-Aided Molecular Design, Vol. 25, Issue 4 https://doi.org/10.1007/s10822-011-9421-5	journal	March 2011
TRPV1 and TRPA1 in cutaneous neurogenic and chronic inflammation: pro-inflammatory response induced by their activation and their sensitization Gouin, Olivier; L’Herondelle, Killian; Lebonvallet, Nicolas Protein & Cell, Vol. 8, Issue 9 https://doi.org/10.1007/s13238-017-0395-5	journal	March 2017
Ruthenium red blocks the capsaicin-induced increase in intracellular calcium and activation of membrane currents in sensory neurones as well as the activation of peripheral nociceptors in vitro Dray, A.; Forbes, C. A.; Burgess, G. M. Neuroscience Letters, Vol. 110, Issue 1-2 https://doi.org/10.1016/0304-3940(90)90786-9	journal	March 1990
A new generation of Ca2+ indicators with greatly improved fluorescence properties. Grynkiewicz, G.; Poenie, M.; Tsien, R. Y. Journal of Biological Chemistry, Vol. 260, Issue 6 https://doi.org/10.1016/S0021-9258(19)83641-4	journal	March 1985
Cloning and functional characterization of the guinea pig vanilloid receptor 1 Savidge, Jonathan; Davis, Clare; Shah, Kirti Neuropharmacology, Vol. 43, Issue 3 https://doi.org/10.1016/S0028-3908(02)00122-3	journal	September 2002
Different responses to repeated applications of zingerone in behavioral studies, recordings from intact and cultured TG neurons, and from VR1 receptors Liu, L.; Welch, J. M.; Erickson, R. P. Physiology & Behavior, Vol. 69, Issue 1-2 https://doi.org/10.1016/S0031-9384(00)00200-6	journal	April 2000
Molecular Basis for Species-Specific Sensitivity to “Hot” Chili Peppers Jordt, Sven-Eric; Julius, David Cell, Vol. 108, Issue 3 https://doi.org/10.1016/S0092-8674(02)00637-2	journal	February 2002
Cloning and functional expression of a human orthologue of rat vanilloid receptor-1 Hayes, Philip; Meadows, Helen J.; Gunthorpe, Martin J. Pain, Vol. 88, Issue 2 https://doi.org/10.1016/S0304-3959(00)00353-5	journal	January 2000
The Cloned Capsaicin Receptor Integrates Multiple Pain-Producing Stimuli Tominaga, Makoto; Caterina, Michael J.; Malmberg, Annika B. Neuron, Vol. 21, Issue 3 https://doi.org/10.1016/S0896-6273(00)80564-4	journal	September 1998
Gαi Controls the Gating of the G Protein-Activated K+ Channel, GIRK Peleg, Sagit; Varon, Dalia; Ivanina, Tatiana Neuron, Vol. 33, Issue 1 https://doi.org/10.1016/S0896-6273(01)00567-0	journal	January 2002
Oxytocin Modulates Nociception as an Agonist of Pain-Sensing TRPV1 Nersesyan, Yelena; Demirkhanyan, Lusine; Cabezas-Bratesco, Deny Cell Reports, Vol. 21, Issue 6 https://doi.org/10.1016/j.celrep.2017.10.063	journal	November 2017
Cloning and pharmacological characterization of mouse TRPV1 Correll, Craig C.; Phelps, P. Tara; Anthes, John C. Neuroscience Letters, Vol. 370, Issue 1 https://doi.org/10.1016/j.neulet.2004.07.058	journal	November 2004
The Ankyrin Repeats of TRPV1 Bind Multiple Ligands and Modulate Channel Sensitivity Lishko, Polina V.; Procko, Erik; Jin, Xiangshu Neuron, Vol. 54, Issue 6 https://doi.org/10.1016/j.neuron.2007.05.027	journal	June 2007
Resiniferatoxin Binds to the Capsaicin Receptor (TRPV1) near the Extracellular Side of the S4 Transmembrane Domain Chou, Margaret Z.; Mtui, Tecla; Gao, Ying-Duo Biochemistry, Vol. 43, Issue 9 https://doi.org/10.1021/bi035981h	journal	March 2004
Structural Insight into Tetrameric hTRPV1 from Homology Modeling, Molecular Docking, Molecular Dynamics Simulation, Virtual Screening, and Bioassay Validations Feng, Zhiwei; Pearce, Larry V.; Xu, Xiaomeng Journal of Chemical Information and Modeling, Vol. 55, Issue 3 https://doi.org/10.1021/ci5007189	journal	February 2015
Nonivamide, a Constituent of Capsicum oleoresin Constant, Howard L.; Cordell, Geoffrey A.; West, Dennis P. Journal of Natural Products, Vol. 59, Issue 4 https://doi.org/10.1021/np9600816	journal	January 1996
Directed deterrence by capsaicin in chillies Tewksbury, Joshua J.; Nabhan, Gary P. Nature, Vol. 412, Issue 6845 https://doi.org/10.1038/35086653	journal	July 2001
The capsaicin receptor: a heat-activated ion channel in the pain pathway Caterina, Michael J.; Schumacher, Mark A.; Tominaga, Makoto Nature, Vol. 389, Issue 6653 https://doi.org/10.1038/39807	journal	October 1997
Molecular basis of infrared detection by snakes Gracheva, Elena O.; Ingolia, Nicholas T.; Kelly, Yvonne M. Nature, Vol. 464, Issue 7291 https://doi.org/10.1038/nature08943	journal	March 2010
TRPV1 structures in distinct conformations reveal activation mechanisms Cao, Erhu; Liao, Maofu; Cheng, Yifan Nature, Vol. 504, Issue 7478 https://doi.org/10.1038/nature12823	journal	December 2013
TRPV1 structures in nanodiscs reveal mechanisms of ligand and lipid action Gao, Yuan; Cao, Erhu; Julius, David Nature, Vol. 534, Issue 7607 https://doi.org/10.1038/nature17964	journal	May 2016
Structural mechanism underlying capsaicin binding and activation of the TRPV1 ion channel Yang, Fan; Xiao, Xian; Cheng, Wei Nature Chemical Biology, Vol. 11, Issue 7 https://doi.org/10.1038/nchembio.1835	journal	June 2015
The vanilloid receptor TRPV1: 10 years from channel cloning to antagonist proof-of-concept Szallasi, Arpad; Cortright, Daniel N.; Blum, Charles A. Nature Reviews Drug Discovery, Vol. 6, Issue 5 https://doi.org/10.1038/nrd2280	journal	May 2007
TRPV1 temperature activation is specifically sensitive to strong decreases in amino acid hydrophobicity Sosa-Pagán, Jason O.; Iversen, Edwin S.; Grandl, Jörg Scientific Reports, Vol. 7, Issue 1 https://doi.org/10.1038/s41598-017-00636-4	journal	April 2017
Effects of piperine, the pungent component of black pepper, at the human vanilloid receptor (TRPV1) McNamara, Fergal N.; Randall, Andrew; Gunthorpe, Martin J. British Journal of Pharmacology, Vol. 144, Issue 6 https://doi.org/10.1038/sj.bjp.0706040	journal	March 2005
Oxidative challenges sensitize the capsaicin receptor by covalent cysteine modification Chuang, H. -h.; Lin, S. Proceedings of the National Academy of Sciences, Vol. 106, Issue 47 https://doi.org/10.1073/pnas.0902675106	journal	November 2009
Acid potentiation of the capsaicin receptor determined by a key extracellular site Jordt, S. -E.; Tominaga, M.; Julius, D. Proceedings of the National Academy of Sciences, Vol. 97, Issue 14 https://doi.org/10.1073/pnas.100129497	journal	June 2000
Activity-dependent targeting of TRPV1 with a pore-permeating capsaicin analog Li, H.; Wang, S.; Chuang, A. Y. Proceedings of the National Academy of Sciences, Vol. 108, Issue 20 https://doi.org/10.1073/pnas.1018550108	journal	May 2011
Low-cost functional plasticity of TRPV1 supports heat tolerance in squirrels and camels Laursen, Willem J.; Schneider, Eve R.; Merriman, Dana K. Proceedings of the National Academy of Sciences, Vol. 113, Issue 40 https://doi.org/10.1073/pnas.1604269113	journal	September 2016
The Endoplasmic Reticulum of Dorsal Root Ganglion Neurons Contains Functional TRPV1 Channels Gallego-Sandín, Sonia; Rodríguez-García, Arancha; Alonso, María Teresa Journal of Biological Chemistry, Vol. 284, Issue 47 https://doi.org/10.1074/jbc.M109.019687	journal	September 2009
C-terminal Dimerization Activates the Nociceptive Transduction Channel Transient Receptor Potential Vanilloid 1 Wang, Shu; Chuang, Huai-hu Journal of Biological Chemistry, Vol. 286, Issue 47 https://doi.org/10.1074/jbc.M111.256669	journal	September 2011
Molecular Cloning and Functional Characterization of Xenopus tropicalis Frog Transient Receptor Potential Vanilloid 1 Reveal Its Functional Evolution for Heat, Acid, and Capsaicin Sensitivities in Terrestrial Vertebrates Ohkita, Masashi; Saito, Shigeru; Imagawa, Toshiaki Journal of Biological Chemistry, Vol. 287, Issue 4 https://doi.org/10.1074/jbc.M111.305698	journal	November 2011
Evolution of Heat Sensors Drove Shifts in Thermosensation between Xenopus Species Adapted to Different Thermal Niches Saito, Shigeru; Ohkita, Masashi; Saito, Claire T. Journal of Biological Chemistry, Vol. 291, Issue 21 https://doi.org/10.1074/jbc.M115.702498	journal	March 2016
Molecular Determinants of Vanilloid Sensitivity in TRPV1 Gavva, Narender R.; Klionsky, Lana; Qu, Yusheng Journal of Biological Chemistry, Vol. 279, Issue 19 https://doi.org/10.1074/jbc.M312577200	journal	March 2004
Activation of transient receptor potential vanilloid 1 (TRPV1) by resiniferatoxin: Activation of TRPV1 by resiniferatoxin Raisinghani, Manish; Pabbidi, Reddy M.; Premkumar, Louis S. The Journal of Physiology, Vol. 567, Issue 3 https://doi.org/10.1113/jphysiol.2005.087874	journal	September 2005
The Cloned Rat Vanilloid Receptor VR1 Mediates Both R-Type Binding and C-Type Calcium Response in Dorsal Root Ganglion Neurons Szallasi, Arpad; Blumberg, Peter M.; Annicelli, Lauren L. Molecular Pharmacology, Vol. 56, Issue 3 https://doi.org/10.1124/mol.56.3.581	journal	September 1999
Similarities and differences in the currents activated by capsaicin, piperine, and zingerone in rat trigeminal ganglion cells Liu, L.; Simon, S. A. Journal of Neurophysiology, Vol. 76, Issue 3 https://doi.org/10.1152/jn.1996.76.3.1858	journal	September 1996
Molecular mechanism of the tree shrew’s insensitivity to spiciness Han, Yalan; Li, Bowen; Yin, Ting-Ting PLOS Biology, Vol. 16, Issue 7 https://doi.org/10.1371/journal.pbio.2004921	journal	July 2018
Detailed Analysis of the Binding Mode of Vanilloids to Transient Receptor Potential Vanilloid Type I (TRPV1) by a Mutational and Computational Study Ohbuchi, Katsuya; Mori, Yoshikazu; Ogawa, Kazuo PLOS ONE, Vol. 11, Issue 9 https://doi.org/10.1371/journal.pone.0162543	journal	September 2016
Capsaicin-induced ion fluxes in dorsal root ganglion cells in culture Wood, Jn; Winter, J.; James, If The Journal of Neuroscience, Vol. 8, Issue 9 https://doi.org/10.1523/JNEUROSCI.08-09-03208.1988	journal	September 1988
Capsaicin Binds to the Intracellular Domain of the Capsaicin-Activated Ion Channel Jung, Jooyoung; Hwang, Sun Wook; Kwak, Jiyeon The Journal of Neuroscience, Vol. 19, Issue 2 https://doi.org/10.1523/JNEUROSCI.19-02-00529.1999	journal	January 1999
Functional Role of C-Terminal Cytoplasmic Tail of Rat Vanilloid Receptor 1 Vlachová, Viktorie; Teisinger, Jan; Sušánková, Klára The Journal of Neuroscience, Vol. 23, Issue 4 https://doi.org/10.1523/JNEUROSCI.23-04-01340.2003	journal	February 2003
The Zebrafish Ortholog of TRPV1 Is Required for Heat-Induced Locomotion Gau, P.; Poon, J.; Ufret-Vincenty, C. Journal of Neuroscience, Vol. 33, Issue 12 https://doi.org/10.1523/jneurosci.5403-12.2013	journal	March 2013
TRPV1: A Potential Drug Target for Treating Various Diseases Brito, Rafael; Sheth, Sandeep; Mukherjea, Debashree Cells, Vol. 3, Issue 2 https://doi.org/10.3390/cells3020517	journal	May 2014
An external sodium ion binding site controls allosteric gating in TRPV1 channels Jara-Oseguera, Andres; Bae, Chanhyung; Swartz, Kenton J. eLife, Vol. 5 https://doi.org/10.7554/eLife.13356	journal	February 2016