1. Structural Biology and Molecular Biophysics
  2. Neuroscience
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Engineering vanilloid-sensitivity into the rat TRPV2 channel

  1. Feng Zhang
  2. Sonya M Hanson
  3. Andres Jara-Oseguera
  4. Dmitriy Krepkiy
  5. Chanhyung Bae
  6. Larry V Pearce
  7. Peter M Blumberg
  8. Simon Newstead
  9. Kenton J Swartz  Is a corresponding author
  1. National Institutes of Health, United States
  2. Memorial Sloan Kettering Cancer Center, United States
  3. National Cancer Institute, United States
  4. University of Oxford, United Kingdom
Research Article
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Cite this article as: eLife 2016;5:e16409 doi: 10.7554/eLife.16409

Abstract

The TRPV1 channel is a detector of noxious stimuli, including heat, acidosis, vanilloid compounds and lipids. The gating mechanisms of the related TRPV2 channel are poorly understood because selective high affinity ligands are not available, and the threshold for heat activation is extremely high (> 50 {degree sign}C). Cryo-EM structures of TRPV1 and TRPV2 reveal that they adopt similar structures, and identify a putative vanilloid binding pocket near the internal side of TRPV1. Here we use biochemical and electrophysiological approaches to investigate the resiniferatoxin (RTx) binding site in TRPV1 and to explore the functional relationships between TRPV1 and TRPV2. Collectively, our results support the interaction of vanilloids with the proposed RTx binding pocket, and demonstrate an allosteric influence of a tarantula toxin on vanilloid binding. Moreover, we show that sensitivity to RTx can be engineered into TRPV2, demonstrating that the gating and permeation properties of this channel are similar to TRPV1.

Article and author information

Author details

  1. Feng Zhang

    Molecular Physiology and Biophysics Section, Porter Neuroscience Research Center, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, United States
    Competing interests
    No competing interests declared.
  2. Sonya M Hanson

    Computational Biology Program, Memorial Sloan Kettering Cancer Center, Bethesda, United States
    Competing interests
    No competing interests declared.
  3. Andres Jara-Oseguera

    Molecular Physiology and Biophysics Section, Porter Neuroscience Research Center, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, United States
    Competing interests
    No competing interests declared.
  4. Dmitriy Krepkiy

    Molecular Physiology and Biophysics Section, Porter Neuroscience Research Center, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, United States
    Competing interests
    No competing interests declared.
  5. Chanhyung Bae

    Molecular Physiology and Biophysics Section, Porter Neuroscience Research Center, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, United States
    Competing interests
    No competing interests declared.
  6. Larry V Pearce

    Laboratory of Cancer Biology and Genetics, Center for Cancer Research, National Cancer Institute, Bethesda, United States
    Competing interests
    No competing interests declared.
  7. Peter M Blumberg

    Laboratory of Cancer Biology and Genetics, Center for Cancer Research, National Cancer Institute, Bethesda, United States
    Competing interests
    No competing interests declared.
  8. Simon Newstead

    Department of Biochemistry, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
  9. Kenton J Swartz

    Molecular Physiology and Biophysics Section, Porter Neuroscience Research Center, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, United States
    For correspondence
    swartzk@ninds.nih.gov
    Competing interests
    Kenton J Swartz, Reviewing editor, eLife.

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocol (#1253-15) of the National Institute of Neurological Disorders and Stroke.

Reviewing Editor

  1. Indira M Raman, Northwestern University, United States

Publication history

  1. Received: March 26, 2016
  2. Accepted: May 12, 2016
  3. Accepted Manuscript published: May 13, 2016 (version 1)
  4. Version of Record published: June 14, 2016 (version 2)

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

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