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This new, fully revised and expanded edition of Ionic Channels of Excitable Membranes includes new chapters on fast chemical synapses, modulation through G protein coupled receptors and second messenger systems, molecules cloning, site directed mutagenesis, and cell biology. It begins with the classical biophysical work of Hodgkin and Huxley and then weaves a description of the known ionic channels together with their biological functions. The book continues by developing the physical and molecular principles needed for explaining permeation, gating, pharmacological modification, and molecular diversity, and ends with a discussion of channel evolution. Ionic Channels of Excitable Membranes is written to be accessible and interesting to biological and physical scientists of all kinds.
This book deals with recent breakthroughs in ion-channel research that have been brought about by the combined effort of experimental biophysicists and computational physicists, who together are beginning to unravel the story of these exquisitely designed biomolecules. With chapters by leading experts, the book is aimed at researchers in nanodevices and biosensors, as well as advanced undergraduate and graduate students in biology and the physical sciences.
A wealth of information has been accumulated about the function of ion channels of excitable cells since the extensive and pioneering voltage clamp studies by Hodgkin, Huxley, and Katz 36 years ago. The study of ion chan nels has now reached a stage at which a quantum jump in progress is antici pated. There are many good reasons for this. Patch clamp techniques origi nally developed by Neher and Sakmann 12 years ago have made it possible to study the function of ion channels in a variety of cells. Membrane ionic currents can now be recorded practically from many types of cells using the whole-cell patch clamp technique. The opening and closing of individual ion channels can be analyzed using the single-channel patch clamp method. Techniques have also been developed to incorporate purified ion channels into lipid bilayers to reconstitute an "excitable membrane. " Advanced tech niques developed in molecular biology, genetics, and immunology, such as gene cloning and the use of monoclonal antibodies, are now being applied to the study of ion channels. A variety of drugs have now been found or are suspected to interact with ion channels to exert therapeutic effects. In addition to the classical exam ples, as represented by local anesthetics, many other drugs, including cal cium antagonists, psychoactive drugs, cardiac drugs, and anticonvulsants, shown to alter ion channel function. For certain pesticides such as have been pyrethroids and DDT, sodium channels are clearly the major target site.
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.
In the past few years, the scientific community has witnessed rapid and significant progress in the study of ion channels. Technological advance ment in biophysics, molecular biology, and immunology has been greatly accelerated, making it possible to conduct experiments that were deemed very difficult if not impossible in the past. For example, patch-clamp tech niques can now be used to measure ionic currents generated by almost any type of cell, thereby allowing us to analyze single-channel events. It is now possible to incorporate purified ion channel components into lipid bilayers to reconstitute an "excitable membrane." Gene cloning and monoclonal antibody techniques provide us with new approaches to the study of the molecular structure of ion channels. A variety of drugs have now been found or are suspected to interact with ion channels to exert therapeutic effects. In addition to the classical exam ples, as represented by local anesthetics, many other drugs, including cal cium antagonists, psychoactive drugs, cardiac drugs, and anticonvulsants, have been shown to alter the ion channel function. For certain pesticides such as pyrethroids and DDT, sodium channels are clearly the major target site. Many diseases of excitable tissues are known to be associated with, if not caused by, dysfunction of ion channels; these include cardiac ar rhythmias, angina pectoris, cystic fibrosis, myotonia, and epilepsies, to men tion only a few. Channel dysfunction can now be studied due to theoretical and technological developments in this area.
This book deals with recent breakthroughs in ion-channel research that have been brought about by the combined effort of experimental biophysicists and computational physicists, who together are beginning to unravel the story of these exquisitely designed biomolecules. With chapters by leading experts, the book is aimed at researchers in nanodevices and biosensors, as well as advanced undergraduate and graduate students in biology and the physical sciences.
Because of the highly significant and widely recognized roles of ion channels in physiology, pathophysiology, pharmacology, and toxicology, the term ion channel has now become a household word in the biomedical sciences. This volume covers preparations and techniques for the study of various ion channels. Both voltage-gated and ligand-gated ion channels of neurons, axons, and cardiac and smooth muscles are covered. It includes not only patch clamp techniques but molecular biology and imaging techniques as well. Key Features * Comprehensive protocols included for the study of: * Ion channels using patch-clamp, molecular biology, and imaging techniques * Role of ion channels in physiology, pathophysiology, pharmacology, and toxicology * Specific ion channels of specific tissues
Ion Channels, Part C, Volume 653 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on a variety of topics, including Nonsense suppression in ion channels, Engineering Ion Channels Using Protein Trans-splicing, Probing Ion Channel Neighborhoods Using APEX, STX based probes for NaVs, ANAP: a versatile, fluorescent probe of ion channel gating and regulation, High Throughput Screens for Small Molecule Ion Channel Modulators, Using toxins to study ion channels, Re/de-constructing ubiquitin regulation of ion channels, Tethered Peptide Toxins for Ion Channels, Voltage-Sensing Phosphatase Molecular Engineering, and more. Additional chapters cover Engineering excitable cells, Stretch and Poke Stimulation of Mechanically-Activated Ion Channels, Optical Control of STIM Channels, High Throughput Electrophysiological Evaluation of Mutant Ion Channels, Evaluating BEST1 Mutations in RPE Stem Cells, Long Read Transcript Profiling of Ion Channel Splice Variants, Permeation of Connexin Channels, Ratiometric pH indicator for melanosomes and lysosomes, and Ion channels in the epithelial cells of the choroid plexus. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology series