Essential Ion Channel Methods

1st Edition - June 14, 2010

Editor: P. Michael Conn

eBook ISBN:

9 7 8 - 0 - 1 2 - 3 8 2 2 3 7 - 6

The rapid growth of interest and research activity in ion channels is indicative of their fundamental importance in the maintenance of the living state. This volume was prepared… Read more

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The rapid growth of interest and research activity in ion channels is indicative of their fundamental importance in the maintenance of the living state. This volume was prepared with a view toward providing a sampling of the range of molecular and physical methods that are significant for the study of ion channels.

As part of the Reliable Lab Solutions series, Essential Ion Channel Methods brings together chapters from volumes 293 and 294 of Methods in Enzymology. The chapters have been selected by the editor and updated, when possible, by their original authors to include new research and references. The result is a set of chapters which make use of graphics, comparisons to other methods, and provide tricks and approaches that make it possible to adapt methods to other systems. Methods are presented in a fashion that allows their replication by individuals new to the field, yet providing valuable information for seasoned investigators.

PART I. Assembly

1. Assembly of Ion Channels
ZuFang Sheng and Carol Deutsch

PART II. Genetics

2. Identification of Ion Channel-Associated Proteins Using the Yeast Two-Hybrid System
Martin Niethammer and Morgan Sheng

PART III. Electrophysiology

3. Patch Clamp Studies of the Cystic Fibrosis Transmembrane Conductance Regulator Chloride Channel
John W. Hanrahan, Zie Kone , Ceri J. Mathews,Jiexin Luo, Yanlin Jia and Paul Lindsell

4. Tight-Seal Whole-cell Patch Clamping of C. elegans Neurons
S.R. Lockery and M.B. Goodman

5. Gating Currents
Enrico Stefani and Francisco Bezanilla

6. Determining Ion Channel Permeation Properties
Ted Begenisich

PART IV. Expression Systems

7. Expression of Ligand-Gated Ion Channels Using Using Semliki Forest Virus and Baculovirus
Kathryn Radford and Gary Buell

8. Recombinant Adenovirus-Mediated Expression in Nervous System of Genes Coding for Ion Channels and Other Molecules Involved in Synaptic Function
Markus U. Ehrengruber, Markus Lanzrein, Youfeng Xu, Mark C. Jasek, David B. Kantor, Erin M. Schuman, Henry A. Lester, and Norman Davidson

9. Transient Expression of Heteromeric Ion Channels
Alison L. Eertmoed, Yolanda F. Vallejo, and William N. Green

PART V. Model Simulations

10. Computer Simulations and Modeling of Ion Channels
Michael E. Green

PART VI. Physical

11. Fluorescence Techniques for Measuring Ion Channel Activity
Gönül Velicelebi, Kenneth A. Stauderman, Mark A. Varney, Michael Akong, Stephen D. Hess, and Edwin C. Johnson

12. Ligand Binding Methods for Analysis of Ion Channel Structure and Function
Steen E. Pedersen, Monica M. Lurtz, and Rao V. L. Papineni

13. Three-dimensional structure of membrane proteins determined by two-dimensional crystallization, electron cryomicroscopy, and image analysis
Mark Yeager, Vinzenz M. Unger, Alok K. Mitra

14. Voltage Clamp Biosensors for Capillary Electrophoresis
Owe Orwar, Kent Jardemark, Cecilia Farre, Ingemar Jacobson, Alexander Moscho, Jason B. Shear, Harvey A. Fishman, Sheri J. Lillard, Richard N. Zare

15. Ion Channels As Tools to Monitor Lipid Bilayer-Membrane Protein Interactions: Gramicidin Channels As Molecular Force
O. S. Andersen, C. Nielsen, A. M. Maer, J. A. Lundbæk, M. Goulian, and R. E. Koeppe I

PART VII. Purification and Reconstitution

16. Purification and Reconstitution of the Epithelial Chloride Channel Cystic Fibrosis Transmembrane Conductance Regulator
Mohabir Ramjeesingh, Elizabeth Garani, Kevin Galley, Canhui Li, Yanchun Wang, Christine E. Bear

17. Reconstitution of Native and Cloned Channels into Planar Bilayers
I. Favre, Y.-M. Sun, and E. Moczydlowski

PART VIII. Second Messengers and Biochemical Approaches

18. Protein Phosphorylation of Ligand-Gated Ion Channels
Andrew l. Mammen, Sunjeev Kamboj, and Richard L. Huganir

19. Analysis of Ion Channel Associated Proteins
Michael Wyszynski and Morgan Sheng

20. Secondary Messenger Regulation of Ion Channels/Plant Patch Clamping
Sarah M. Assmann and Lisa Romano

PART IX. Special Channels

21. ATP-Sensitive Potassium Channels
Joseph Bryan, M. Schwanstecher, C. Schwanstecher, F. Chudziak, U. Panten, J.P. Clement IV, G. Gonzalez, and L. Aguilar-Bryan

22. Simplified Fast Pressure-Clamp Technique for Studying Mechanically-Gated Channels
Don W. McBride, Jr. and Owen P. Hamill

23. Purification and Heterologous Expression of Inhibitory Glycine Receptors
Bodo Laube and Heinrich Betz

24. Functional Analyses of Aquaporin Water Channel Proteins
Peter Agre, John C. Mathai, Barbara L. Smith, and Gregory M. Preston

PART X. Toxins and Other Membrane Active Compounds

25. Conus Peptides as Probes for Ion Channels
J. Michael McIntosh, Baldomero M. Olivera, and Lourdes J. Cruz

26. Scorpion Toxins as Tools for Studying Potassium Channels
Maria Garcia, Markus Hanner, Hans-Günther Knaus, Robert Slaughter, Gregory J. Kaczorowski

27. The Use of Planar Lipid Bilayer Membranes for Rapid Screening of Membrane Active Compounds
Bruce L. Kagan, Tajb A. Mirzabekov, Antoly Y. Silberstein

28. Antibodies to Ion Channels
Angela Vincent, Ian Hart, Ashwin Pinto, and F. Anne Stephenson

P. Michael Conn

P. Michael Conn is the Senior Vice President for Research and Associate Provost, Texas Tech Health Sciences Center. He is The Robert C. Kimbrough, Professor of Internal Medicine and Cell Biology/Biochemistry. He was previously Director of Research Advocacy and Professor of Physiology and Pharmacology, Cell Biology and Development and Obstetrics and Gynecology at Oregon Health and Science University and Senior Scientist of the Oregon National Primate Research Center (ONPRC). He served for twelve years as Special Assistant to the President and Associate Director of the ONPRC. After receiving a B.S. degree and teaching certification from the University of Michigan (1971), a M.S. from North Carolina State University (1973), and a Ph.D. degree from Baylor College of Medicine (1976), Conn did a fellowship at the NIH, then joined the faculty in the Department of Pharmacology, Duke University Medical Center where he was promoted to Associate Professor in 1982. In 1984, he became Professor and Head of Pharmacology at the University of Iowa College of Medicine, a position he held for eleven years. Conn is known for his research in the area of the cellular and molecular basis of action of gonadotropin releasing hormone action in the pituitary and therapeutic approaches that restore misfolded proteins to function. His work has led to drugs that have benefitted humans and animals. Most recently, he has identified a new class of drugs, pharmacoperones, which act by regulating the intracellular trafficking of receptors, enzymes and ion channels. He has authored or co-authored over 350 publications in this area and written or edited over 200 books, including texts in neurosciences, molecular biology and endocrinology. Conn has served as the editor of many professional journals and book series (Endocrinology, Journal of Clinical Endocrinology and Metabolism, Endocrine, Methods, Progress in Molecular Biology and Translational Science and Contemporary Endocrinology). Conn served on the National Board of Medical Examiners, including two years as chairman of the reproduction and endocrinology committee. The work of his laboratory has been recognized with a MERIT award from the NIH, the J.J. Abel Award of the American Society for Pharmacology and Experimental Therapeutics, the Weitzman, Oppenheimer and Ingbar Awards of the Endocrine Society, the National Science Medal of Mexico (the Miguel Aleman Prize) and the Stevenson Award of Canada. He is the recipient of the Oregon State Award for Discovery, the Media Award of the American College of Neuropsychopharmacology and was named a distinguished Alumnus of Baylor College of Medicine in 2012. Conn is a previous member of Council for the American Society for Cell Biology and the Endocrine Society and is a prior President of the Endocrine Society, during which time he founded the Hormone Foundation and worked with political leadership to heighten the public’s awareness of diabetes. Conn’s students and fellows have gone on to become leaders in industry and academia. He is an elected member of the Mexican Institute of Medicine and a fellow of the American Association for the Advancement of Science. He is the co-author of The Animal Research War (2008) and many articles for the public and academic community on the value of animal research and the dangers posed by animal extremism. His op/eds have appeared in The Washington Post, The LA Times, The Wall Street Journal, the Des Moines Register, and elsewhere. Conn consults with organizations that are influenced by animal extremism and with universities and companies facing challenges from these groups.

Affiliations and expertise

Senior Vice President for Research and Associate Provost, Texas Tech Health Sciences Center, TX, USA