Protocol

Ionotropic Glutamate Receptor Technologies

Volume 106 of the series Neuromethods pp 83-100

Expression, Purification, and Crystallization of Full Length Ionotropic Glutamate Receptors

  • Maria V. YelshanskayaAffiliated withDepartment of Biochemistry and Molecular Biophysics, Columbia University
  • , Kei SaotomeAffiliated withDepartment of Biochemistry and Molecular Biophysics, Columbia University
  • , Minfen LiAffiliated withDepartment of Biochemistry and Molecular Biophysics, Columbia University
  • , Alexander I. SobolevskyAffiliated withDepartment of Biochemistry and Molecular Biophysics, Columbia University Email author 

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Abstract

Ionotropic glutamate receptors (iGluRs) are ligand-gated ion channels that mediate the majority of excitatory neurotransmission in brain and are implicated in nearly all aspects of central nervous system development and function. There is great clinical interest in iGluRs because their dysfunction is associated with chronic neurodegenerative conditions, psychiatric disorders, brain trauma and stroke. Despite physiological and pathophysiological importance, obtaining structural information about full-length iGluRs has proven to be a difficult task. To facilitate further structural/functional studies, we describe a detailed protocol of the full-length iGluR expression, purification, and crystallization.

Key words

Ionotropic glutamate receptor Membrane protein Ion channel Structure Insect cells Protein expression Protein purification Baculovirus X-ray crystallography Diffraction Crystals