Hyperpolarizing vasodilators activate ATP-sensitive K+ channels in arterial smooth muscle

Vasodilators are used clinically for the treatment of hypertension and heart failure. The effects of some vasodilators seem to be mediated by membrane hyperpolarization. The molecular basis of this hyperpolarization has been investigated by examining the properties of single K+ channels in arterial smooth muscle cells. The presence of adenosine triphosphate (ATP)-sensitive K+ channels in these cells was demonstrated at the single channel level. These channels were opened by the hyperpolarizing vasodilator cromakalim and inhibited by the ATP-sensitive K+ channel blocker glibenclamide. Furthermore, in arterial rings the vasorelaxing actions of the drugs diazoxide, cromakalim, and pinacidil and the hyperpolarizing actions of vasoactive intestinal polypeptide and acetylcholine were blocked by inhibitors of the ATP-sensitive K+ channels, suggesting that all these agents may act through a common pathway in smooth muscle by opening ATP-sensitive K+ channels.     

Membrane-Potential Alteration During K＋ Uptake of Different Salt-Tolerant Wheat Varieties

K＋ is the most abundant cation in plant cells and plays an important role in many ways.K＋ uptake of plant has respect to its salt resistant capacity.There are two categories of channel transportation for plants to uptake K＋,one is through K＋ channels and the other is through nonselective cation channels（NSCCs）.The transmembrane localization of K＋ may change membrane potential（MP）.In this paper,three wheat varieties with different salt tolerance were selected and the MP was measured by microelectrode during K＋ uptake.The results showed that the effects of K＋ uptake on MP through K＋ channels or NSCCs were distinct.K＋ influx through K＋ channels led to MP hyperpolarization,while K＋ influx through NSCCs resulted in depolarization.Diverse MP alteration of wheat varieties with different salt tolerance was mainly due to NSCCs-mediated K＋ uptake.Compared with the salt-tolerant wheat,the MP hyperpolarization during K＋ uptake of saltsensitive wheat was much more evident,probably because of the cation outflux through NSCCs during this process.     

Protein kinase activity closely associated with a reconstituted calcium-activated potassium channel.

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.     

Protective role of ATP-sensitive potassium channels in hypoxia-induced generalized seizure

Adenosine triphosphate (ATP)-sensitive potassium (K(ATP)) channels are activated by various metabolic stresses, including hypoxia. The substantia nigra pars reticulata (SNr), the area with the highest expression of K(ATP) channels in the brain, plays a pivotal role in the control of seizures. Mutant mice lacking the Kir6.2 subunit of K(ATP) channels [knockout (KO) mice] were susceptible to generalized seizures after brief hypoxia. In normal mice, SNr neuron activity was inactivated during hypoxia by the opening of the postsynaptic K(ATP) channels, whereas in KO mice, the activity of these neurons was enhanced. K(ATP) channels exert a depressant effect on SNr neuronal activity during hypoxia and may be involved in the nigral protection mechanism against generalized seizures.     

胞外ATP、H2O2、Ca2+与NO调控木榄根系离子平衡机理研究

运用非损伤微测技术(NMT),研究了短期盐胁迫下胞外ATP(eATP)、H2 O2 、Ca2 + 与NO 对非泌盐红树木榄根 系K+/Na+ 平衡的调控作用。NaCl(100 mmol/L,24 h)与等渗甘露醇处理的实验表明,木榄根尖对盐胁迫的响应具 有高度的离子特异性。盐胁迫增强了木榄根尖的Na+ 外流,但Na+ 外流被Na+ /H+ 逆向转运蛋白抑制剂Amiloride 和质膜H+ -ATPase 抑制剂Vanadate 抑制,表明Na+ 外流源于根尖表皮细胞质膜Na+ /H+ 逆向转运系统驱动的Na+ 外排。短期盐胁迫处理能诱导木榄根尖K+ 外流,但被氯化四乙胺(TEA,外向K+ 通道抑制剂)明显抑制,证明K+ 外流是由激活的去极化外向型离子通道KORCs 介导。胞外ATP(300 mol/L)、H2 O2 (10 mmol/L)、Ca2 + (10 mmol/ L)与SNP(NO 供体,100 mol/L)均能增加短期盐胁迫下的Na+ 外流,同时抑制K+ 外流。其中,促进Na+ 外流效果 较强的是H2 O2 和Ca2 + ,而Ca2 + 和NO 抑制K+ 外流的效果突出。这些实验结果表明,胞外ATP、H2 O2 、Ca2 + 与NO 这4 种盐胁迫信使是通过上调木榄根系细胞质膜Na+ /H+ 逆向转运体系(Na+ /H+ 逆向转运体和H+ 泵)活性,在促 进Na+ 和H+ 逆向跨膜转运的同时,抑制去极化激活的K+ 离子通道来减少盐诱导的K+ 外流。      

Defense strategies against hypoxia and hypothermia

Because aerobic metabolic rates decrease in hypoxia-sensitive cells under oxygen-limiting conditions, the demand for glucose or glycogen for anaerobic glycolysis may rise drastically as a means of making up for the energetic shortfall. However, ion and electrical potentials typically cannot be sustained because of energy insufficiency and high membrane permeabilities; therefore metabolic and membrane functions in effect become decoupled. In hypoxia-tolerant animals, these problems are resolved through a number of biochemical and physiological mechanisms; of these metabolic arrest and stabilized membrane functions are the most effective strategies for extending tolerance to hypoxia. Metabolic arrest is achieved by means of a reversed or negative Pasteur effect (reduced or unchanging glycolytic flux at reduced O2 availability); and coupling of metabolic and membrane function is achievable, in spite of the lower energy turnover rates, by maintaining membranes of low permeability (probably via reduced densities of ion-specific channels). The possibility of combining metabolic arrest with channel arrest has been recognized as an intervention strategy. To date, the success of this strategy has been minimal, mainly because depression of metabolism through cold is the usual arrest mechanism used, and hypothermia in itself perturbs controlled cell function in most endotherms.     

过表达胡杨PeRIN4基因拟南芥提高质膜H+-ATPase活性和耐盐性

本文克隆了RIN4(RPM1-interacting protein 4)在胡杨中的同源基因PeRIN4,并在拟南芥中进行过表达,通过研究转基因株系的耐盐表型、质膜H+-ATPsae活性及H+ 、Na+、K+等的动态离子流,揭示了PeRIN4基因在植物响应和适应盐胁迫环境中的作用。利用定位载体pGreen0029-PeRIN4-GFP瞬时转化拟南芥叶肉细胞原生质体的方法,对胡杨PeRIN4蛋白进行亚细胞定位,发现该蛋白定位在细胞的胞质中。耐盐表型实验结果显示,在100 mmol/L NaCl处理下,拟南芥PeRIN4过表达株系(OE1和OE8)的生存率和根长均明显高于野生型(WT)和转空载体拟南芥(VC),说明PeRIN4基因能够提高拟南芥的耐盐性。与WT和VC相比,拟南芥PeRIN4过表达株系质膜H+-ATPsae的活性较高。动态离子流数据显示,在盐胁迫下,PeRIN4过表达株系外排H+和Na+ 离子的能力强于野生型和转空载体拟南芥,然而K+的外流却弱于WT和VC。因此,PeRIN4蛋白具有调节质膜H+-ATPsae活性的功能。拟南芥质膜H+-ATPsae活性的提高主要有两方面的作用:一是可以增强H+泵的质子动力势,驱动Na+/H+逆向转运蛋白,提高Na+外排的能力;二是抑制质膜的去极化,减少K+离子通过去极化激活的外向型K+通道(DA-KORCs)和非选择性阳离子通道(DA-NSCCs)外流,维持了K+/Na+平衡,从而提高PeRIN4转基因拟南芥的耐盐性。      

NMDA receptor-mediated K+ efflux and neuronal apoptosis

Neuronal death induced by activating N-methyl-D-aspartate (NMDA) receptors has been linked to Ca2+ and Na+ influx through associated channels. Whole-cell recording from cultured mouse cortical neurons revealed a NMDA-evoked outward current, INMDA-K, carried by K+ efflux at membrane potentials positive to -86 millivolts. Cortical neurons exposed to NMDA in medium containing reduced Na+ and Ca2+ (as found in ischemic brain tissue) lost substantial intracellular K+ and underwent apoptosis. Both K+ loss and apoptosis were attenuated by increasing extracellular K+, even when voltage-gated Ca2+ channels were blocked. Thus NMDA receptor-mediated K+ efflux may contribute to neuronal apoptosis after brain ischemia.     

Ion selectivity in a semisynthetic K+ channel locked in the conductive conformation

Potassium channels are K+-selective protein pores in cell membrane. The selectivity filter is the functional unit that allows K+ channels to distinguish potassium (K+) and sodium (Na+) ions. The filter's structure depends on whether K+ or Na+ ions are bound inside it. We synthesized a K+ channel containing the d-enantiomer of alanine in place of a conserved glycine and found by x-ray crystallography that its filter maintains the K+ (conductive) structure in the presence of Na+ and very low concentrations of K+. This channel conducts Na+ in the absence of K+ but not in the presence of K+. These findings demonstrate that the ability of the channel to adapt its structure differently to K+ and Na+ is a fundamental aspect of ion selectivity, as is the ability of multiple K+ ions to compete effectively with Na+ for the conductive filter.     

Cytoprotective role of Ca2+- activated K+ channels in the cardiac inner mitochondrial membrane

Ion channels on the mitochondrial inner membrane influence cell function in specific ways that can be detrimental or beneficial to cell survival. At least one type of potassium (K+) channel, the mitochondrial adenosine triphosphate-sensitive K+ channel (mitoKATP), is an important effector of protection against necrotic and apoptotic cell injury after ischemia. Here another channel with properties similar to the surface membrane calcium-activated K+ channel was found on the mitochondrial inner membrane (mitoKCa) of guinea pig ventricular cells. MitoKCa significantly contributed to mitochondrial K+ uptake of the myocyte, and an opener of mitoKCa protected hearts against infarction.     

Crystal structure of a mammalian voltage-dependent Shaker family K+ channel

Voltage-dependent potassium ion (K+) channels (Kv channels) conduct K+ ions across the cell membrane in response to changes in the membrane voltage, thereby regulating neuronal excitability by modulating the shape and frequency of action potentials. Here we report the crystal structure, at a resolution of 2.9 angstroms, of a mammalian Kv channel, Kv1.2, which is a member of the Shaker K+ channel family. This structure is in complex with an oxido-reductase beta subunit of the kind that can regulate mammalian Kv channels in their native cell environment. The activation gate of the pore is open. Large side portals communicate between the pore and the cytoplasm. Electrostatic properties of the side portals and positions of the T1 domain and beta subunit are consistent with electrophysiological studies of inactivation gating and with the possibility of K+ channel regulation by the beta subunit.     

Turnover, membrane insertion, and degradation of sodium channels in rabbit urinary bladder

Noise analysis of rabbit bladder revealed two components: Lorentzian noise, arising from interaction of amiloride with the Na+ channel, and flicker noise (l/f, where f is frequency), as in other biological membranes. Hydrostatic pressure, which causes exchange between intracellular vesicular membrane and apical membrane, increases the number but not the single-channel current of the amiloride-sensitive channels. Flicker noise arises from degraded channels that have lost amiloride sensitivity and Na+ to K+ selectivity. The degraded channels were selectively removed by washing the mucosal surface. These results imply channel turnover by intracellular synthesis, transfer from vesicular to apical membrane, degradation, and elimination.     

alpha-Klotho as a regulator of calcium homeostasis

alpha-klotho was identified as a gene associated with premature aging-like phenotypes characterized by short lifespan. In mice, we found the molecular association of alpha-Klotho (alpha-Kl) and Na+,K+-adenosine triphosphatase (Na+,K+-ATPase) and provide evidence for an increase of abundance of Na+,K+-ATPase at the plasma membrane. Low concentrations of extracellular free calcium ([Ca2+]e) rapidly induce regulated parathyroid hormone (PTH) secretion in an alpha-Kl- and Na+,K+-ATPase-dependent manner. The increased Na+ gradient created by Na+,K+-ATPase activity might drive the transepithelial transport of Ca2+ in cooperation with ion channels and transporters in the choroid plexus and the kidney. Our findings reveal fundamental roles of alpha-Kl in the regulation of calcium metabolism.     

K+及主要离子间交互作用对凡纳滨对虾存活、生长及体内ATP酶的影响

采取L16(215)正交设计试验,开展5周凡纳滨对虾养殖试验,分析养殖水体中K+及K+、盐度、Ca2+、Mg2+四因子间交互作用对凡纳滨对虾存活、生长及体内ATP酶的影响。结果表明:K+对凡纳滨对虾成活率、体长和体重增加及体内ATP酶活性均具有显著影响,K+浓度为150 mg/L时成活率最高,而K+浓度为50 mg/L时生长速度最快,Na+-K+-ATPase、Mg2+-ATPase、Ca2+-ATPase酶活也最高;K+、盐度、Ca2+、Mg2+四因子间交互作用对凡纳滨对虾存活、生长及体内ATP酶也具有显著影响。本试验因素水平组合中凡纳滨对虾成活率的最佳组合为盐度15、K+150 mg/L、Ca2+100 mg/L、Mg2+300 mg/L,生长速度的最佳组合为盐度15、K+50 mg/L、Ca2+100 mg/L、Mg2+100 mg/L,凡纳滨对虾体内Na+-K+-ATPase、Mg2+-ATPase、Ca2+-ATPase酶活的最佳组合为盐度5、K+50 mg/L、Ca2+100 mg/L、Mg2+100 mg/L。     

Voltage-dependent calcium channels in glial cells

The electrophysiological properties of glial cells were examined in primary culture in the presence of tetraethylammonium and Ba2+, a treatment that reduces K+ permeability of the membrane and enhances currents through voltage-dependent Ca2+ channels. Under these conditions, glial cells showed both spontaneous action potentials and action potentials evoked by the injections of current. These responses appear to represent entry of Ba2+ through Ca2+ channels because they were resistant to tetrodotoxin but were blocked by Mn2+ or Cd2+.     

Sequence of a probable potassium channel component encoded at Shaker locus of Drosophila

Potassium currents are crucial for the repolarization of electrically excitable membranes, a role that makes potassium channels a target for physiological modifications that alter synaptic efficacy. The Shaker locus of Drosophila is thought to encode a K+ channel. The sequence of two complementary DNA clones from the Shaker locus is reported here. The sequence predicts an integral membrane protein of 70,200 daltons containing seven potential membrane-spanning sequences. In addition, the predicted protein is homologous to the vertebrate sodium channel in a region previously proposed to be involved in the voltage-dependent activation of the Na+ channel. These results support the hypothesis that Shaker encodes a structural component of a voltage-dependent K+ channel and suggest a conserved mechanism for voltage activation.     

Studies on the regulation of fatty acid and cholesterol synthesis in avian liver

Homogenates of pigeon liver have been incubated with acetate-C(14) in excess. Simultaneously, substrate for glycolysis was provided as glucose-6-phosphate. The incorporation of carbon-14 into cholesterol was maximal at low levels of glycolysis, whereas fatty acid turnover was maximum at higher glycolytic levels. A regulatory mechanism is proposed to explain the differential synthesis of cholesterol and fatty acids.     

Membrane insertion of a potassium-channel voltage sensor

The mechanism of voltage gating in K+ channels is controversial. The paddle model posits that highly charged voltage-sensor domains move relatively freely across the lipid bilayer in response to membrane depolarization; competing models picture the charged S4 voltage-sensor helix as being shielded from lipid contact by other parts of the protein. We measured the apparent free energy of membrane insertion of a K+-channel S4 helix into the endoplasmic reticulum membrane and conclude that S4 is poised very near the threshold of efficient bilayer insertion. Our results suggest that the paddle model is not inconsistent with the high charge content of S4.     

葡萄糖酸钠对光滑球拟酵母CCTCC M202019能量代谢和丙酮酸积累的影响

在好氧条件下,光滑球拟酵母(Torulopsis glabrata)CCTCC M202019能够利用葡萄糖酸钠为唯一碳源生长并积累少量丙酮酸。在含有90 g/L葡萄糖的培养基中,16 h时添加10 g/L葡萄糖酸钠,与100 g/L葡萄糖作为唯一碳源的对照组相比,细胞质量浓度、葡萄糖消耗速率、丙酮酸生产强度分别提高7.96%、13.04%和32.81%;其原因在于葡萄糖酸钠的添加,使胞内NAD+浓度提高了29.73%,NADH/NAD+比值和ATP含量分别下降26.89%和8.50%,此变化显著缓解了NADH和ATP对于糖酵解途径的抑制作用,从而显著加快糖酵解速率。