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Related Experiment Videos

Subunit arrangement in P2X receptors.

Lin-Hua Jiang1, Miran Kim, Valeria Spelta

  • 1Institute of Molecular Physiology, University of Sheffield, Sheffield S10 2TN, United Kingdom.

The Journal of Neuroscience : the Official Journal of the Society for Neuroscience
|October 3, 2003
PubMed
Summary

Researchers mapped the structure of ATP-gated ionotropic receptors (P2X receptors) using disulfide bonds. This revealed a "head-to-tail" arrangement of subunits in P2X2/3 receptors, crucial for nerve sensation.

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Biophysics

Background:

  • P2X receptors are ATP-gated ion channels vital for nervous system functions, including sensory perception.
  • A key example is the P2X2/3 receptor, involved in primary afferent sensation.
  • Understanding the quaternary structure of these receptors is essential for elucidating their function.

Purpose of the Study:

  • To investigate the subunit arrangement within the quaternary structure of P2X receptors.
  • To determine the proximity of transmembrane domains in P2X receptor subunits.
  • To elucidate the specific subunit composition and arrangement in hetero-oligomeric P2X2/3 receptors.

Main Methods:

  • Engineered cysteine mutations in P2X2 and P2X4 receptor subunits to form disulfide bonds.

Related Experiment Videos

  • Expression of modified subunits in HEK 293 cells.
  • Assessing channel function via ATP-evoked currents and the requirement for dithiothreitol reduction.

    • Main Results:

    • Disulfide bond formation confirmed close proximity between the outer ends of the first and second transmembrane domains of adjacent subunits.
    • Functional assays demonstrated that disulfide bond formation, and thus channel opening, required prior reduction.
    • Analysis of hetero-oligomeric P2X2/3 receptors indicated adjacent P2X3 subunits but not adjacent P2X2 subunits.

    Conclusions:

    • The findings suggest a "head-to-tail" arrangement of subunits in the quaternary structure of P2X receptors.
    • A trimeric P2X2/3 receptor is proposed to have a P2X2(P2X3)2 composition.
    • This structural insight advances our understanding of P2X receptor assembly and function in sensory pathways.