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Coarse Architecture of the Transient Receptor Potential Vanilloid 1 (TRPV1) Ion Channel Determined by Fluorescence Resonance Energy Transfer
Authors:Víctor De-la-Rosa  Gisela E. Rangel-Yescas  Ernesto Ladrón-de-Guevara  Tamara Rosenbaum  León D. Islas
Affiliation:From the Departamento de Fisiología, Facultad de Medicina, and ;§Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Mexico City, D.F., C.P. 04510, Mexico
Abstract:
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 the N or C terminus 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.
Keywords:Imaging   Pain   Patch Clamp   Plasma Membrane   Spectroscopy   TRP Channels
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