细胞功能测定

One of the most important functions of the kidney is the filtration of the blood that takes place in the glomeruli. Glomerular epithelial cells (podocytes) have several functions, including regulation of the filtration process and glomerular basement membrane turnover. Dysfunction ...

Ion channels play key roles in physiology. They function as protein transducers able to transform stimuli and chemical gradients into electrical signals. They also are critical for cell signaling and play a particularly important role in epithelial transport acting as gateways for the m ...

Mature human or mouse erythrocytes functionally express an unexpected diversity of ion channels that endows these small enucleated cells with a toolkit for electrosignaling. Being largely dormant under resting conditions, these ion channels enable erythrocytes to quickly res ...

Electrophysiological recordings from an acutely sliced preparation provide information on ionic currents and excitability of native neurons under near physiological conditions. Although this technique is commonly used on central nervous system structures such as spinal ...

Dendrites emerging from the cell bodies of neurons receive the majority of synaptic inputs. They possess a plethora of ion channels that are essential for the processing of these synaptic signals. To fully understand how dendritic ion channels influence neuronal information processi ...

As large, multimeric, integral membrane proteins, ion channels pose technical challenges to analysis by NMR spectroscopy. Here we present a strategy to overcome some of these technical hurdles, using a representative ion channel modulatory domain, the regulator of K+ conductance (RCK) ...

Regulator of K+ conductance (RCK) domains form a conserved class of ligand-binding domains that control the activity of a variety of prokaryotic and eukaryotic K+ channels. Structural analysis of these domains by X-ray crystallography has provided insight toward mechanisms underly ...

Cysteine contains a highly reactive thiol group and therefore under oxidizing conditions a disulfide bond can form between a pair of cysteines that are juxtaposed in the close vicinity, which can be only reversed by reducing agents. These attributes have been elegantly exploited to study the ...

Ion channels mediate a wide variety of physiological processes by forming small pores across the membranes that allow regulated flow of ions into or out of the cell. The primary linear sequences of ion channel proteins, like any proteins, are composed by 20 different amino acids, each of which is dete ...

There is demand for isoform-specific ion channel inhibitors as tools to investigate the biology of �endogenous ion channels and validate them as targets in drug discovery programs. There is also hope that such inhibitors may be new therapeutic agents or provide the foundation for such agent ...

This chapter describes immunochemistry-based methods to investigate recycling of membrane proteins at the cell surface. Two methods are described, one qualitative and the other quantitative. Both methods consist of two rounds of extracellular antibody capture. Firstly, a prima ...

Steady-state fluorescence spectroscopy is a biophysical technique widely employed to characterize �interactions between proteins in vitro. Only a few proteins naturally fluoresce in cells, but by covalently attaching fluorophores virtually all proteins can be monitored. O ...

Understanding the molecular mechanisms of protein–protein interactions at the cell surface of living cells is fundamental to identifying the nature of cellular processes. Here, we discuss how fluorescence-based approaches have been successfully developed to visualize prot ...

Ion channels are integral membrane proteins that allow the flow of ions across membranes down their electrochemical gradients and are a major determinant of cellular excitability. They play an important role in a variety of biological processes as diverse as insulin release from beta cel ...

The excised inside-out patch clamp technique gives rapid access to the intracellular surface of the plasma membrane while measuring channel activity. This way the effects of intracellular regulators of ion channels or transporters can be studied in isolation, in the absence of most of the c ...

Planar lipid bilayer is an electrophysiological technique that enables study of functional activities of ion channels, porins, and other pore-forming molecular complexes. The main purpose of this method is to monitor ion channels’ behavior at the single molecule level in the artifici ...

The patch clamp technique revolutionized the study of ion channels and is considered the gold standard of measuring ion channel activity, from the academic laboratory to industrial-scale drug screening. This technique enables the study of ion channels, from single molecules up to the who ...

Two-electrode voltage clamp (TEVC) is a conventional electrophysiological technique used to artificially control the membrane potential (V m) of large cells to study the properties of electrogenic membrane proteins, especially ion channels. It makes use of two intracellular ele ...

Microdomains that form on the plasma membrane of cells are essential for signalling compartmentation within cells. The localization of ion channels in these surface microdomains is important in defining what signalling cascades will be generated. For example, in cardiomyocytes, s ...

Ion channels are integral membrane proteins that regulate the flow of ions across the plasma membrane and the membranes of intracellular organelles of both excitable and non-excitable cells. Ion channels are vital to a wide variety of biological processes and are prominent components of t ...