Titlebook: Ion Channels; Methods and Protocol James D. Stockand,Mark S. Shapiro Book 2006 Humana Press 2006 Escherichia coli.Lipid.Vivo.molecular biol

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Biophysical Approach to Determine the Subunit Stoichiometry of the Epithelial Sodium Channel Using thods to determine the functional subunit stoichiometry of ENaC based on a biophysical approach that was first introduced in 1991 to determine the subunit stoichiometry of a voltage-gated K. channel. The strategy is to analyze channel sensitivity to a specific blocker when various mixtures of block-s

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Spectroscopy-Based Quantitative Fluorescence Resonance Energy Transfer AnalysisA key to successful application of green fluorescent protein-based FRET is to reliably separate the FRET signal from various non-FRET fluorescence emissions that coexist in any experimental system. This chapter introduces a FRET quantification method that is based on fluorescence spectroscopic micro

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Probing the Effects of Phosphoinositides on Ion Channels. Membrane phospho-lipids containing phosphoinositides have recently emerged as important regulators of many ion channels, including inward rectifier K . channel, voltage-gated K . and Ca . channels, transient receptor potential channels, and intracellular inositol-1,4,5-trisphos-phate receptor ion

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Epithelial Sodium Channel in Planar Lipid Bilayersodium handling, they serve diverse functions in many tissues. Improper functioning of ENaC has been implicated in several diseases, including salt-sensitive hypertension (Liddle’s syndrome), salt-wasting syndrome (pseudohypoaldosteronism type I), pulmonary edema, and cystic fibrosis. We have utilize

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A Simple In Vivo Method for Assessing Changes of Membrane-Bound Ion Channel Density in , Oocytesd transporters. In the case of ion channels, activity can be easily measured by conventional two-electrode voltage clamping. However, this method only measures the sum of the activity of all plasma membrane-bound channels. Therefore, this measurement cannot discriminate between effects on channel de

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Preparation of Cortical Brain Slices for Electrophysiological Recordingnd whole-cell function in juvenile and adult tissue. Ion channel recordings from small dendritic branches axons in brain slices have demonstrated considerable functional differences in ion channel function within subregions of single cells. These findings have greatly increased understanding of neur

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Carbon Fiber Amperometry in the Study of Ion Channels and Secretion. signals, the induction of exocytosis—the process in which the contents of secretory vesicles are released by their fusion to the plasma membrane—is one of the most important. For example, in neurons and endocrine cells, it allows intercellular communication and secretion of biomolecules. Exocytosi

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KCNQ1 K+ Channel—Mediated Cardiac Channelopathiesciates with different β-subunits of the KCNE protein family. In the human heart, KCNQ1 associates with KCNE1 to generate the .. current characterized by its slow activation and deactivation kinetics. Mutations in either KCNQ1 or KCNE1 are responsible for at least four channelopathies that lead to ca