Ion Channels
The diverse applications in this volume range from the study of allosteric regulation of ion channel activity using a classic mutagenesis approach, to the study of channel subunit stoichiometry using a novel biophysical approach based on fluorescence resonance energy transfer. Highlights include methods for heterologous expression of ion channels in cells, for determining channel structure-function, and for studying channel regulation. It is now clear that ion channels play essential roles in cell biology and physiology and that their dysfunction is the root cause of many human diseases. In Ion Channels: Methods and Protocols, internationally recognized biomedical scientists describe recent technological breakthroughs and demonstrate their use in successful
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The diverse applications in this volume range from the study of allosteric regulation of ion channel activity using a classic mutagenesis approach, to the study of channel subunit stoichiometry using a novel biophysical approach based on fluorescence resonance energy transfer. Highlights include methods for heterologous expression of ion channels in cells, for determining channel structure-function, and for studying channel regulation. It is now clear that ion channels play essential roles in cell biology and physiology and that their dysfunction is the root cause of many human diseases. In Ion Channels: Methods and Protocols, internationally recognized biomedical scientists describe recent technological breakthroughs and demonstrate their use in successful experimental designs. The diverse applications range from the study of allosteric regulation of ion channel activity using a classic mutagenesis approach, to the study of channel subunit stoichiometry using a novel biophysical approach based on fluorescence resonance energy transfer. Highlights include methods for heterologous expression of ion channels in cells, for determining channel structure-function, for studying channel regulation and physiological function, and for genetic screening and investigating channelopathies. The protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and highly practical, Ion Channels: Methods and Protocols offer both novice and experienced researchers a versatile collection of proven techniques for illuminating the structure, function, and regulation of ion channel proteins and finding suitable drug targets when they cause disease.Part I. Methods for Exogenous Expression of Ion Channels in Cells Functional Reconstitution of the Human Epithelial Na+ Channel in a Mammalian Expression System Alexander Staruschenko, Rachell E. Booth, Oleh Pochynyuk, James D. Stockand, and Qiusheng Tong Overexpression of Proteins in Neurons Using Replication-Deficient Virus Richard M. Ahlquist and Jane M. Sullivan Exogenous Expression of Proteins in Neurons Using the Biolistic Particle Delivery System Nikita Gamper and Mark S. Shapiro Part II. Methods for Studying Channel Structure-Function Tertiary and Quaternary Structure Formation of Voltage-Gated Potassium Channels John M. Robinson, Andrey Kosolapov, and Carol Deutsch Biophysical Approach to Determine the Subunit Stoichiometry of the Epithelial Sodium Channel Using the Xenopus laevis Oocyte Expression System Farhad Kosari, Shaohu Sheng, and Thomas R. Kleyman Spectroscopy-Based Quantitative Fluorescence Resonance Energy Transfer Analysis Jie Zheng Part III. Methods for Studying Channel Regulation and Physiological Function Probing the Effects of Phosphoinositides on Ion Channels Chou-Long Huang Epithelial Sodium Channel in Planar Lipid Bilayers Bakhrom K. Berdiev and Dale J. Benos A Simple In Vivo Method for Assessing Changes of Membrane- Bound Ion Channel Density in Xenopus Oocytes Mouhamed S. Awayda, Weijian Shao, Ivana Vukojicic, and Abderrahmane Bengrine Preparation of Cortical Brain Slices for Electrophysiological Recording Costa M. Colbert Juxtacellular Labeling and Chemical Phenotyping of Extracellularly Recorded Neurons In Vivo Glenn M. Toney and Lynette C. Daws Carbon Fiber Amperometry in the Study of Ion Channels and Secretion Duk-Su Koh Part IV. Methods for Studying Channelopathies, Genetic Screening, and Molecular Biology Genetic Screening for Functionality of Bacterial Potassium Channel Mutants Using K+ Uptake-Deficient Escherichia coli Lyubov V. Parfenova and Brad S. Rothberg KCNQ1 K+ Channel-Mediated Cardiac Channelopathies Gildas Loussouarn, Isabelle Baró, and Denis Escande Tissue-Specific Transgenic and Knockout Mice Andrée Porret, Anne-Marie Mérillat, Sabrina Guichard, Friedrich Beermann, and Edith Hummler Index
Comprehensive and highly practical, Ion Channels: Methods and Protocols offer both novice and experienced researchers a versatile collection of proven techniques for illuminating the structure, function, and regulation of ion channel proteins and finding suitable drug targets when they cause disease.Part I. Methods for Exogenous Expression of Ion Channels in Cells Functional Reconstitution of the Human Epithelial Na+ Channel in a Mammalian Expression System Alexander Staruschenko, Rachell E. Booth, Oleh Pochynyuk, James D. Stockand, and Qiusheng Tong Overexpression of Proteins in Neurons Using Replication-Deficient Virus Richard M. Ahlquist and Jane M. Sullivan Exogenous Expression of Proteins in Neurons Using the Biolistic Particle Delivery System Nikita Gamper and Mark S. Shapiro Part II. Methods for Studying Channel Structure-Function Tertiary and Quaternary Structure Formation of Voltage-Gated Potassium Channels John M. Robinson, Andrey Kosolapov, and Carol Deutsch Biophysical Approach to Determine the Subunit Stoichiometry of the Epithelial Sodium Channel Using the Xenopus laevis Oocyte Expression System Farhad Kosari, Shaohu Sheng, and Thomas R. Kleyman Spectroscopy-Based Quantitative Fluorescence Resonance Energy Transfer Analysis Jie Zheng Part III. Methods for Studying Channel Regulation and Physiological Function Probing the Effects of Phosphoinositides on Ion Channels Chou-Long Huang Epithelial Sodium Channel in Planar Lipid Bilayers Bakhrom K. Berdiev and Dale J. Benos A Simple In Vivo Method for Assessing Changes of Membrane- Bound Ion Channel Density in Xenopus Oocytes Mouhamed S. Awayda, Weijian Shao, Ivana Vukojicic, and Abderrahmane Bengrine Preparation of Cortical Brain Slices for Electrophysiological Recording Costa M. Colbert Juxtacellular Labeling and Chemical Phenotyping of Extracellularly Recorded Neurons In Vivo Glenn M. Toney and Lynette C. Daws Carbon Fiber Amperometry in the Study of Ion Channels and Secretion Duk-Su Koh Part IV. Methods for Studying Channelopathies, Genetic Screening, and Molecular Biology Genetic Screening for Functionality of Bacterial Potassium Channel Mutants Using K+ Uptake-Deficient Escherichia coli Lyubov V. Parfenova and Brad S. Rothberg KCNQ1 K+ Channel-Mediated Cardiac Channelopathies Gildas Loussouarn, Isabelle Baró, and Denis Escande Tissue-Specific Transgenic and Knockout Mice Andrée Porret, Anne-Marie Mérillat, Sabrina Guichard, Friedrich Beermann, and Edith Hummler Index
The diverse applications in this volume range from the study of allosteric regulation of ion channel activity using a classic mutagenesis approach, to the study of channel subunit stoichiometry using a novel biophysical approach based on fluorescence resonance energy transfer. Highlights include methods for heterologous expression of ion channels in cells, for determining channel structure-function, and for studying channel regulation. It is now clear that ion channels play essential roles in cell biology and physiology and that their dysfunction is the root cause of many human diseases. In Ion Channels: Methods and Protocols, internationally recognized biomedical scientists describe recent technological breakthroughs and demonstrate their use in successful