Reconstitution of Endoplasmic Reticulum InsP3 Receptors into Black Lipid Membranes

Ilya Bezprozvanny

doi:10.1101/pdb.prot073106

Reconstitution of Endoplasmic Reticulum InsP₃ Receptors into Black Lipid Membranes

Ilya Bezprozvanny1,2,3

¹Department of Physiology, University of Texas Southwestern Medical Center, Dallas, Texas 75390;
²Laboratory of Molecular Neurodegeneration, St. Petersburg State Polytechnical University, St. Petersburg 195251, Russia

Abstract

Intracellular ion channels, including endoplasmic reticulum (ER) calcium (Ca²⁺) channels, are most often studied through their reconstitution into planar lipid bilayers (also called black lipid membranes, or BLMs). General methods for making bilayers and for ion channel reconstitution into BLMs have been extensively detailed elsewhere; thus, here the focus is on specific details relevant for inositol(1,4,5)-trisphosphate receptor (InsP₃R) recordings. These procedures describe how to perform single-channel recordings of native or recombinant InsP₃Rs in BLMs. Similar procedures are used to study native or recombinant ryanodine receptors (RyanRs) in BLMs.