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1.
In many epithelial cells the chloride conductance of the apical membrane increases during the stimulation of electrolyte secretion. Single-channel recordings from human airway epithelial cells showed that beta-adrenergic stimulation evoked apical membrane chloride channel activity, but this response was absent in cells from patients with cystic fibrosis (CF). However, when membrane patches were excised from CF cells into media containing sufficient free calcium (approximately 180 nanomolar), chloride channels were activated. The chloride channels of CF cells were similar to those of normal cells as judged by their current-voltage relations, ion selectivity, and kinetic behavior. These findings demonstrate the presence of chloride channels in the apical membranes of CF airway cells. Their regulation by calcium appears to be intact, but cyclic adenosine monophosphate (cAMP)-dependent control of their activity is defective. 相似文献
2.
We cloned and characterized a protein kinase and ion channel, TRP-PLIK. As part of the long transient receptor potential channel subfamily implicated in control of cell division, it is a protein that is both an ion channel and a protein kinase. TRP-PLIK phosphorylated itself, displayed a wide tissue distribution, and, when expressed in CHO-K1 cells, constituted a nonselective, calcium-permeant, 105-picosiemen, steeply outwardly rectifying conductance. The zinc finger containing alpha-kinase domain was functional. Inactivation of the kinase activity by site-directed mutagenesis and the channel's dependence on intracellular adenosine triphosphate (ATP) demonstrated that the channel's kinase activity is essential for channel function. 相似文献
3.
Protein kinase activity closely associated with a reconstituted calcium-activated potassium channel. 总被引：14，自引：0，他引：14
S K Chung P H Reinhart B L Martin D Brautigan I B Levitan 《Science (New York, N.Y.)》1991,253(5019):560-562
Modulation of the activity of potassium and other ion channels is an essential feature of nervous system function. The open probability of a large conductance Ca(2+)-activated K+ channel from rat brain, incorporated into planar lipid bilayers, is increased by the addition of adenosine triphosphate (ATP) to the cytoplasmic side of the channel. This modulation takes place without the addition of protein kinase, requires Mg2+, and is mimicked by an ATP analog that serves as a substrate for protein kinases but not by a nonhydrolyzable ATP analog. Addition of protein phosphatase 1 reverses the modulation by MgATP. Thus, there may be an endogenous protein kinase activity firmly associated with this K+ channel. Some ion channels may exist in a complex that contains regulatory protein kinases and phosphatases. 相似文献
4.
导管经历了完整的细胞程序性死亡过程(programmed cell death,PCD),形成运送水分的管状通道;而筛分子分化则经历了细胞程序性半死亡过程,形成运输有机养分的通道。为了进一步弄清二者在小麦(Triticumaestivum L.)颖果发育过程中超微结构上的变化,本研究利用生物电镜和酶的超微细胞化学定位技术比较了筛分子和导管分化过程中超微结构的变化,以及酸性磷酸酶(ACPase,acid phosphatase)活性的分布动态。结果表明:筛分子呈对称性半圆形分布于导管的两侧,且筛分子内细胞核的降解晚于导管,筛分子细胞质主要通过液泡膜内陷包裹细胞器,形成自噬体进行降解,最终仍保留了部分细胞器碎片,筛分子仍然存活;而在导管分化中,主要是液泡破裂后,胞内细胞质被完全降解,导管最终死亡,形成管状分子。ACPase作为液泡的标志性酶,在导管液泡膜破裂后,多定位于线粒体等发生降解的细胞器上。而在筛分子中,不仅在发生降解的线粒体等细胞器上检测到了ACPase活性,在成熟筛分子胞间连丝上也检测到了ACPase活性,表明ACPase不仅参与了小麦颖果筛分子和导管发育中PCD进程,可能还与细胞间物质运输有关。 相似文献
5.
The effects of calcium removal on the voltage-dependent potassium channels of isolated squid neurons were studied with whole cell patch-clamp techniques. When the calcium ion concentration was lowered from 10 to 0 millimolar (that is, no added calcium), potassium channel activity, identified from its characteristic time course, disappeared within a few seconds and there was a parallel increase in resting membrane conductance and in the holding current. The close temporal correlation of the changes in the three parameters suggests that potassium channels lose their ability to close in the absence of calcium and simultaneously lose their selectivity. If potassium channels were blocked by barium ion before calcium ion was removed, the increases in membrane conductance and holding current were delayed or prevented. Thus calcium is an essential cofactor in the gating of potassium channels in squid neurons. 相似文献
6.
Calcium uptake into mitochondria occurs via a recently identified ion channel called the uniporter. Here, we characterize the phylogenomic distribution of the uniporter's membrane-spanning pore subunit (MCU) and regulatory partner (MICU1). Homologs of both components tend to co-occur in all major branches of eukaryotic life, but both have been lost along certain protozoan and fungal lineages. Several bacterial genomes also contain putative MCU homologs that may represent prokaryotic calcium channels. The analyses indicate that the uniporter may have been an early feature of mitochondria. 相似文献
7.
Reconstitution and phosphorylation of chloride channels from airway epithelium membranes 总被引：4，自引：0，他引：4
Airway epithelial chloride secretion is controlled by the apical-membrane chloride permeability. Purified apical-membrane vesicles from bovine tracheal epithelium have now been shown to contain functional chloride channels by using the planar-bilayer technique. Three types of chloride channels were observed; a voltage-dependent, calcium-independent, 71-picoSiemen (in 150 mM NaCl) channel accounted for more than 80 percent of the vesicular chloride conductance and was under strict control of phosphorylation. The channel underwent a fast rundown in less than 2 to 3 minutes of recording, and reactivation required in situ exposure to a phosphorylating "cocktail" containing the catalytic subunit of the adenosine 3',5'-monophosphate (cAMP)-dependent protein kinase. Mean open time and open probability were increased after phosphorylation, whereas slope conductance remained unchanged. Thus, metabolic control of tracheal chloride single channels can now be studied in vitro. 相似文献
8.
Effect of adenosine 3',5'-monophosphate on neuronal pacemaker activity: a voltage clamp analysis 总被引：1，自引：0，他引：1
S N Treistman 《Science (New York, N.Y.)》1981,211(4477):59-61
Bursting pacemaker activity in nerve cells can be modified for long periods by synaptic input of short duration. There is evidence that cyclic nucleotides may play a role in these modifications. The predominant effect of elevated levels of adenosine 3',5'-monophosphate in Aplysia neurons was an increased slope conductance to hyperpolarizing pulses, evident in voltage clamp records. A similar increase in slope conductance was seen as one component of maximum strength synaptic stimulation, which is consistent with the idea that cyclic nucleotides are important in the expression of synaptic alteration of bursting pacemaker activity. 相似文献
9.
Exchange of conduction pathways between two related K+ channels 总被引：26，自引：0，他引：26
H A Hartmann G E Kirsch J A Drewe M Taglialatela R H Joho A M Brown 《Science (New York, N.Y.)》1991,251(4996):942-944
The structure of the ion conduction pathway or pore of voltage-gated ion channels is unknown, although the linker between the membrane spanning segments S5 and S6 has been suggested to form part of the pore in potassium channels. To test whether this region controls potassium channel conduction, a 21-amino acid segment of the S5-S6 linker was transplanted from the voltage-activated potassium channel NGK2 to another potassium channel DRK1, which has very different pore properties. In the resulting chimeric channel, the single channel conductance and blockade by external and internal tetraethylammonium (TEA) ion were characteristic of the donor NGK2 channel. Thus, this 21-amino acid segment controls the essential biophysical properties of the pore and may form the conduction pathway of these potassium channels. 相似文献
10.
Ion channels in yeast 总被引：21，自引：0，他引：21
M C Gustin B Martinac Y Saimi M R Culbertson C Kung 《Science (New York, N.Y.)》1986,233(4769):1195-1197
Voltage-dependent ion channels have been found in the plasma membrane of the yeast Saccharomyces cerevisiae. Ion channel activities were recorded from spheroplasts or patches of plasma membrane with the patch-clamp technique. The most prominent activities came from a set of potassium channels with the properties of activation by positive but not negative voltages, high selectivity for potassium over sodium ion, unit conductance of 20 picosiemens, inhibition by tetraethylammonium or barium ions, and bursting kinetics. 相似文献
11.
Single subunits of the GABAA receptor form ion channels with properties of the native receptor 总被引：13，自引：0，他引：13
L A Blair E S Levitan J Marshall V E Dionne E A Barnard 《Science (New York, N.Y.)》1988,242(4878):577-579
The alpha and beta subunits of the gamma-aminobutyric acidA (GABAA) receptor were expressed individually in Xenopus oocytes by injection of RNA synthesized from their cloned DNAs. GABA-sensitive chloride channels were detected several days after injection with any one of three different alpha RNAs (alpha 1, alpha 2, and alpha 3) or with beta RNA. The channels induced by each of the alpha-subunit RNAs were indistinguishable, they had multiple conductance levels (10, 19, 28, and 42 picosiemens), and their activity was potentiated by pentobarbital and inhibited by picrotoxin. The beta channels usually expressed poorly but showed similar single channel conductance levels (10, 18, 27, and 40 picosiemens), potentiation by pentobarbital and inhibition by picrotoxin. The finding that both alpha and beta subunits, examined separately, form GABA-sensitive ion channels with permeation properties and regulatory sites characteristic of the native receptor suggests that the amino acid sequences that confer these properties are within the homologous domains shared by the subunits. 相似文献
12.
Magnus CJ Lee PH Atasoy D Su HH Looger LL Sternson SM 《Science (New York, N.Y.)》2011,333(6047):1292-1296
Ionic flux mediates essential physiological and behavioral functions in defined cell populations. Cell type-specific activators of diverse ionic conductances are needed for probing these effects. We combined chemistry and protein engineering to enable the systematic creation of a toolbox of ligand-gated ion channels (LGICs) with orthogonal pharmacologic selectivity and divergent functional properties. The LGICs and their small-molecule effectors were able to activate a range of ionic conductances in genetically specified cell types. LGICs constructed for neuronal perturbation could be used to selectively manipulate neuron activity in mammalian brains in vivo. The diversity of ion channel tools accessible from this approach will be useful for examining the relationship between neuronal activity and animal behavior, as well as for cell biological and physiological applications requiring chemical control of ion conductance. 相似文献
13.
An apical-membrane chloride channel in human tracheal epithelium 总被引：11，自引：0，他引：11
M J Welsh 《Science (New York, N.Y.)》1986,232(4758):1648-1650
The mechanism of chloride transport by airway epithelia has been of substantial interest because airway and sweat gland-duct epithelia are chloride-impermeable in cystic fibrosis. The decreased chloride permeability prevents normal secretion by the airway epithelium, thereby interfering with mucociliary clearance and contributing to the morbidity and mortality of the disease. Because chloride secretion depends on and is regulated by chloride conductance in the apical cell membrane, the patch-clamp technique was used to directly examine single-channel currents in primary cultures of human tracheal epithelium. The cells contained an anion-selective channel that was not strongly voltage-gated or regulated by calcium in cell-free patches. The channel was also blocked by analogs of carboxylic acid that decrease apical chloride conductance in intact epithelia. When attached to the cell, the channel was activated by isoproterenol, although the channel was also observed to open spontaneously. However, in some cases, the channel was only observed after the patch was excised from the cell. These results suggest that this channel is responsible for the apical chloride conductance in airway epithelia. 相似文献
14.
Mitochondria and chloroplasts are essential eukaryotic organelles of endosymbiotic origin. Dynamic cellular machineries divide these organelles. The mechanisms by which mitochondria and chloroplasts divide were thought to be fundamentally different because chloroplasts use proteins derived from the ancestral prokaryotic cell division machinery, whereas mitochondria have largely evolved a division apparatus that lacks bacterial cell division components. Recent findings indicate, however, that both types of organelles universally require dynamin-related guanosine triphosphatases to divide. This mechanistic link provides fundamental insights into the molecular events driving the division, and possibly the evolution, of organelles in eukaryotes. 相似文献
15.
Increased activity of calcium leak channels in myotubes of Duchenne human and mdx mouse origin 总被引：25，自引：0，他引：25
Elevated free Ca2+ concentrations found in adult dystrophic muscle fibers result in enhanced protein degradation. Since the difference in concentrations may reflect differences in entry, Ca2+ leak channels in cultures of normal and Duchenne human myotubes, and normal and mdx murine myotubes, have been identified and characterized. The open probability of leak channels is markedly increased in dystrophic myotubes. Other channel properties, such as mean open times, single channel conductance, ion selectivity, and behavior in the presence of pharmacological agents, were similar among myotube types. Compared to the Ca2+ concentrations in normal human and normal mouse myotubes, intracellular resting free Ca2+ concentrations ([Ca2+]i) in myotubes of Duchenne and mdx origin were significantly higher at a time when dystrophin is first expressed in normal tissue. Taken together, these findings suggest that the increased open probability of Ca2+ leak channels contributes to the elevated free intracellular Ca2+ concentration in Duchenne human and mdx mouse myotubes. 相似文献
16.
Calmodulin activation of calcium-dependent sodium channels in excised membrane patches of Paramecium 总被引：8，自引：0，他引：8
Calmodulin is a calcium-binding protein that participates in the transduction of calcium signals. The electric phenotypes of calmodulin mutants of Paramecium have suggested that the protein may regulate some calcium-dependent ion channels. Calcium-dependent sodium single channels in excised patches of the plasma membrane from Paramecium were identified, and their activity was shown to decrease after brief exposure to submicromolar concentrations of calcium. Channel activity was restored to these inactivated patches by adding calmodulin that was isolated from Paramecium to the cytoplasmic surface. This restoration of channel activity did not require adenosine triphosphate and therefore, probably resulted from direct binding of calmodulin, either to the sodium channel itself or to a channel regulator that was associated with the patch membrane. 相似文献
17.
Uptake of isolated chloroplasts by mammalian cells 总被引：3，自引：0，他引：3
M M Nass 《Science (New York, N.Y.)》1969,165(898):1128-1131
Mouse fibroblasts (L cells) in suspension culture incorporated isolated chloroplasts of spinach and African violets and isolated mitochondria of chicken liver. The organelles resided in the cell cytoplasm and were not contained in vacuoles or digestion vesicles. Green cells divided like normal cells. Green chloroplasts were followed for five cell generations or 5 days, at which time hybrid cells were greatly outnumbered by nongreen progeny cells. The ingested chloroplasts retained their structural integrity as determined by electron microscopy of organelles and hybrid cells and by analysis of photochemical activity and DNA in chloroplasts reisolated from cells after 1 or 2 days in culture. 相似文献
18.
BRONK JR 《Science (New York, N.Y.)》1963,141(3583):816-818
Triiodothyronine administered to thyroidectomized rats preferentially increased the total capacity of the electron transport phosphorylation system when isolated mitochondria from the liver were tested with different substrates, but it caused little increase in the activity of the slowest dehydrogenases and no uncoupling. The increased activity appeared to be partly due to direct activation of some component of the electron transport system, although triiodothyronine injection also stimulated the incorporation of amino acids by isolated mitochondria. 相似文献
19.
李凤兰 《北京林业大学学报》1992,14(1):46-50,T001
运用电镜及电镜细胞化学等方法对大波斯菊小孢子发生时期,即小孢子母细胞到四分休的整个发育过程中的线粒体动态进行了观察。实验表明:在小孢子母细胞减数分裂过程中,线粒体要经受一个复杂的反分化过程。在减数分裂前期的早期阶段,线粒体的数量增加很快;与此同时,一些空白的被称作”spherical inclusion”的细胞器(STs)也随之产生,它们代表着新产生细胞器的退化。造成退化的原因可能是由于细胞器缺少核密码的调控。推测减数分裂后线粒体围核现象可能与ATP、信息大分子及其他物质的传递有关。 相似文献
20.
Identification of synaptophysin as a hexameric channel protein of the synaptic vesicle membrane 总被引：24，自引：0，他引：24
L Thomas K Hartung D Langosch H Rehm E Bamberg W W Franke H Betz 《Science (New York, N.Y.)》1988,242(4881):1050-1053
The quaternary structure and functional properties of synaptophysin, a major integral membrane protein of small presynaptic vesicles, were investigated. Cross-linking and sedimentation studies indicate that synaptophysin is a hexameric homo-oligomer, which in electron micrographs exhibits structural features common to channel-forming proteins. On reconstitution into planar lipid bilayers, purified synaptophysin displays voltage-sensitive channel activity with an average conductance of about 150 picosiemens. Because specific channels and fusion pores have been implicated in vesicular uptake and release of secretory compounds, synaptophysin may have a role in these processes. 相似文献