Coarse architecture of the transient receptor potential vanilloid 1 (TRPV1) ion channel determined by fluorescence resonance energy transfer
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The transient receptor potential vanilloid 1 ion channel is responsible for the perception of high temperatures and low extracellular pH, and it is also involved in the response to some pungent compounds. Importantly, it is also associated with the perception of pain and noxious stimuli. Here, we attempt to discern the molecular organization and location of the N and C termini of the transient receptor potential vanilloid 1 ion channel by measuring FRET between genetically attached enhanced yellow and cyan fluorescent protein to theNorCterminus of the channel protein, expressed in transfected HEK 293 cells or Xenopus laevis oocytes. The static measurements of the domain organization were mapped into an available cryo-electron microscopy density of the channel with good agreement. These measurements also provide novel insights into the organization of terminal domains and their proximity to the plasma membrane. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.
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Cryo-electron microscopy; Cyan fluorescent protein; Fluorescence resonance energy transfer; High temperature; Molecular organization; Static measurements; Transient receptor potential vanilloid; Xenopus laevis oocytes; Cell membranes; Electron microscopes; Energy transfer; Enzyme activity; Fluorescence; Proteins; Ions; cyan fluorescent protein; vanilloid receptor 1; yellow fluorescent protein; amino terminal sequence; animal cell; article; carboxy terminal sequence; cell membrane; cellular distribution; controlled study; cryoelectron microscopy; embryo; fluorescence resonance energy transfer; gene expression; genetic organization; genetic transfection; human; human cell; nonhuman; priority journal; protein domain; Imaging; Pain; Patch Clamp; Plasma Membrane; Spectroscopy; TRP Channels; Algorithms; Animals; Cell Membrane; Cryoelectron Microscopy; Female; Fluorescence Resonance Energy Transfer; HEK293 Cells; Humans; Ion Channel Gating; Kinetics; Luminescent Proteins; Membrane Potentials; Models, Molecular; Oocytes; Patch-Clamp Techniques; Protein Multimerization; Protein Structure, Tertiary; Rats; TRPV Cation Channels; Xenopus laevis
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