Ion Channel Reconstitution

ISBN-13: 9781475713633 / Angielski / Miękka / 2013 / 577 str.

C. Miller

Opis

It is now over 30 years since the idea of ion-conducting pores burst on the elec- trophysiological scene, 15 years since these were generalIy realized to be mem- brane-spanning proteins, and 10 years since the first observations of single ion channels from higher organisms were made. During the past 5 years, several integral membrane channel proteins have been purified in a functionalIy competent state: the nicotinic acetylcholine receptor, the Na + channel, mitochondrial "VDAC," and a variety of porins. The stage is thus set to attack ion channels in the same ways that biochemists have been attacking enzymes for decades: isolation folIowed by functional analysis in as simple a system as possible, with a view towards understanding the molecular mechanisms ofthe protein's behavior and how this is related to the underlying molecular structure. This is always a daunting task, alI the more so with ion channels because of our still primitive and scanty understanding of channel structures and because of the difficulty in iso- lating functionally active channel proteins. In this volume, which can be considered a biochemically slanted companion to Sakmann and Neher's Single-Channel Recording, I have tried to present a view of the current landscape of ion-channel reconstitution. These chapters illustrate not only the different approaches and techniques of the major practitioners of ion- channel reconstitution but, as importantly, the varied motivations for doing this kind of work.

Spis treści

I. Basics.- 1 The Physical Nature of Planar Bilayer Membranes.- 2 Ion Channel Electrostatics and the Shapes of Channel Proteins.- 3 Superoxide Dismutase as a Model Ion Channel.- 4 Single-Channel Enzymology.- 5 How to Set Up a Bilayer System.- 6 Fusion of Liposomes to Planar Bilayers.- 7 Incorporation of Ion Channels by Fusion.- II. Nicotinic Acetylcholine Receptor.- 8 The Reconstituted Acetylcholine Receptor.- 9 Immunologic Analysis of the Acetylcholine Receptor.- 10 Function of Acetylcholine Receptors in Reconstituted Liposomes.- III. Sodium Channel.- 11 Skeletal Muscle Sodium Channels: Isolation and Reconstitution.- 12 Reconstitution of the Sodium Channel from Electrophorus electricus.- 13 The Reconstituted Sodium Channel from Brain.- 14 Gating of Batrachotoxin-Activated Sodium Channels in Lipid Bilayers.- 15 Ion Conduction Through Sodium Channels in Planar Lipid Bilayers.- 16 Blocking Pharmacology of Batrachotoxin-Activated Sodium Channels.- IV. Other Channels in Model Membranes.- 17 The Large Calcium-Activated Potassium Channel.- 18 The Sarcoplasmic Reticulum Potassium Channel: Lipid Effects.- 19 Characterization of Dihydropyridine-Sensitive Calcium Channels from Purified Skeletal Muscle Transverse Tubules.- 20 Calcium Channels.- 21 Phosphorylation of a Reconstituted Potassium Channel.- 22 Voltage Gating in VDAC: Toward a Molecular Mechanism.- 23 Analysis and Chemical Modification of Bacterial Porins.