小麦幼苗拒Na+部位的拒Na+机理

采用日立Z 80 0 0原子吸收分光光度计测Na 、K 含量 ,采用不连续蔗糖梯度离心分离质膜和液泡膜微囊。递增盐度和盐冲击处理下 ,耐盐品种德抗 96 1的SK ,Na(吸收 ) 值和SK ,Na(运输 ) 值均明显大于盐敏感品种鲁麦 15 ;德抗 96 1根部和鲁麦 15根茎结合部Na 含量均呈递增趋势 ,表现出累积效应 ;德抗 96 1根细胞质膜微囊和液泡膜微囊H ATP酶活性均明显大于鲁麦15 ,鲁麦 15根茎结合部液泡膜微囊H ATP酶活性大于德抗 96 1,在同一品种的植株里 ,盐冲击的根和根茎结合部细胞质膜微囊和液泡膜微囊H ATP酶活性均小于递增盐度的酶活性。小麦拒Na 部位细胞质膜和液泡膜H ATP酶活性与其耐盐性强弱成正相关     

高等植物Na+ 吸收、转运及细胞内Na+ 稳态平衡研究进展

盐胁迫是影响农业生产的重要环境因素之一。本文对植物Na+吸收的机制和途径、Na+在植物体内的长距离转运以及细胞内Na+稳态平衡的研究进展进行了概述。参与植物Na+吸收与转运的蛋白和通道可能包括HKT、LCT1、AKT和NSCC等。其中, HKT是植物体内普遍存在的一类转运蛋白, 能够介导Na+的吸收, 其结构中的带电氨基酸残基对于其离子选择性有着非常明显的影响。LCT1是从小麦中发现的一类能够介导低亲和性阳离子吸收的蛋白, 然而在典型的土壤Ca2+浓度下LCT1并不能发挥吸收Na+的功能。AKT家族的成员在高盐环境下可能也参与了Na+的吸收。目前虽然还没有克隆到编码NSCC蛋白的基因, 但是NSCC作为植物吸收Na+的主要途径的观点已被广泛接受。SOS1和HKT参与了Na+在根部与植株地上部的长距离转运过程, 它们在木质部和韧皮部的Na+装载和卸载中发挥重要作用, 从而影响植物的抗盐性。另外, 由质膜Na+/H+逆向转运蛋白SOS1、蛋白激酶SOS2以及Ca2+结合蛋白SOS3组成的SOS复合体对细胞的Na+稳态具有重要的调节作用, 单子叶和双子叶植物之间的这种调节机制在结构和功能上具有保守性。SOS复合体与其它位于质膜或液泡膜上的Na+/H+逆向转运蛋白以及H+泵一起调节着细胞的Na+稳态。     

高等植物Na+吸收、转运及细胞内Na+稳态平衡研究进展

盐胁迫是影响农业生产的重要环境因素之一。本文对植物Na 吸收的机制和途径、Na 在植物体内的长距离转运以及细胞内Na 稳态平衡的研究进展进行了概述。参与植物Na 吸收与转运的蛋白和通道可能包括HKT、LCT1、AKT和NSCC等。其中,HKT是植物体内普遍存在的一类转运蛋白,能够介导Na 的吸收,其结构中的带电氨基酸残基对于其离子选择性有着非常明显的影响。LCT1是从小麦中发现的一类能够介导低亲和性阳离子吸收的蛋白,然而在典型的土壤Ca2 浓度下LCT1并不能发挥吸收Na 的功能。AKT家族的成员在高盐环境下可能也参与了Na 的吸收。目前虽然还没有克隆到编码NSCC蛋白的基因,但是NSCC作为植物吸收Na 的主要途径的观点已被广泛接受。SOS1和HKT参与了Na 在根部与植株地上部的长距离转运过程,它们在木质部和韧皮部的Na 装载和卸载中发挥重要作用,从而影响植物的抗盐性。另外,由质膜Na /H 逆向转运蛋白SOS1、蛋白激酶SOS2以及Ca2 结合蛋白SOS3组成的SOS复合体对细胞的Na 稳态具有重要的调节作用,单子叶和双子叶植物之间的这种调节机制在结构和功能上具有保守性。SOS复合体与其它位于质膜或液泡膜上的Na /H 逆向转运蛋白以及H 泵一起调节着细胞的Na 稳态。     

Export of Na+ from cells of the halotolerant microalga Dunaliella maritima: Na+/H+ antiporter or primary Na+-pump?

Transport of Na+ and K+ ions through the plasma membrane of intact cells of the halotolerant microalga Dunaliella maritima Massjuk was studied. Ion fluxes through the plasma membrane were induced by hyperosmotic shock (uptake of Na+ by the cells is transformed into extrusion of Na+) or by addition of K+ to a suspension of K+-deficient cells (uptake of K+ by the cells is associated with extrusion of Na+). The pathway of Na+ extrusion from the D. maritima cells does not depend on the direction or value of the proton gradient on the plasma membrane. In particular, the efficiency of Na+ extrusion was not changed at extracellular pH values varying from 6.0 to 8.0. The protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP) (20 microM) and the H+-ATPase inhibitor N,N-dicyclohexyl carbodiimide (DCCD) (25 and 100 microM) inhibited accumulation of K+ by the cells but did not influence Na+ extrusion. Significant acidification of the medium did not induce a net current of Na+ from the cells through a Na+/H+ antiporter. The data indicate that the Na+/H+ antiporter of the plasma membrane of D. maritima is not responsible for Na+ extrusion from the cells. These results can be explained by the involvement of a primary electrogenic Na+ pump (a Na+-transporting ATPase) in Na+ transfer through the plasma membrane of this alga.     

The sided action of Na+ and of K+ on reconstituted shark (Na+ + K+)-ATPase engaged in Na+−Na+ exchange accompanied by ATP hydrolysis. I. The ATP activation curve

The ATP hydrolysis dependent Na⁺-Na⁺ exchange of reconstituted shark (Na⁺ + K⁺)-ATPase is electrogenic with a transport stoichiometry as for the Na⁺-K⁺ exchange, suggesting that translocation of extracellular Na⁺ is taking place via the same route as extracellular K⁺. The preparation thus offers an opportunity to compare the sided action of Na⁺ and of K⁺ on the affinity for ATP in a reaction in which the intermediary steps in the overall reaction seems to be the same without and with K⁺. With Na⁺ but no K⁺ on the two sides of the enzyme, the ATP-activation curve is hyperbolic and the affinity for ATP is high. Extracellular K⁺ in concentrations of 50 μM (the lowest tested) and up gives biphasic ATP activation curves, with both a high- and a low-affinity component for ATP. Cytoplasmic K⁺ also gives biphasic ATP-activation curves, however, only when the K⁺ concentration is 50 mM or higher (Na⁺ + K⁺ = 130 mM). The different ATP-activation curves are explained from the Albers-Post scheme, in which there is an ATP-dependent and an ATP-independent deocclusion of E₂(Na₂⁺) and E₂(K₂⁺), respectively, and in which the dephosphorylation of E₂-P is rate limiting in the presence of Na⁺ (but no K⁺) extracellular, whereas in the presence of extracellular K⁺ it is the deocclusion of E₂(K₂⁺) which is rate limiting.     

HCO 3 − -coupled Na+ influx is a major determinant of Na+ turnover and Na+/K+ pump activity in rat hepatocytes

Summary Recent studies in hepatocytes indicate that Na⁺-coupled HCO ₃ ^– transport contributes importantly, to regulation of intracellular pH and membrane HCO ₃ ^– transport. However, the direction of net coupled Na⁺ and HCO ₃ ^– movement and the effect of HCO ₃ ^– on Na⁺ turnover and Na⁺/K⁺ pump activity are not known. In these studies, the effect of HCO ₃ ^– on Na⁺ influx and turnover were measured in primary rat hepatocyte cultures with²²Na⁺, and [Na⁺] _i was measured in single hepatocytes using the Na⁺-sensitive fluorochrome SBFI. Na⁺/K⁺ pump activity was measured in intact perfused rat liver and hepatocyte monolayers as Na⁺-dependent or ouabain-suppressible⁸⁶Rb uptake, and was measured in single hepatocytes as the effect of transient pump inhibition by removal of extracellular K⁺ on membrane potential difference (PD) and [Na⁺] _i . In hepatocyte monolayers, HCO ₃ ^– increased²²Na⁺ entry and turnover rates by 50–65%, without measurably altering²²Na⁺ pool size or cell volume, and HCO ₃ ^– also increased Na⁺/K⁺ pump activity by 70%. In single cells, exposure to HCO ₃ ^– produced an abrupt and sustained rise in [Na⁺] _i , from 8 to 12mm. Na⁺/K⁺ pump activity assessed in single cells by PD excursions during transient K⁺ removal increased 2.5-fold in the presence of HCO ₃ ^– , and the rise in [Na⁺] _i produced by inhibition of the Na⁺/K⁺ pump was similarly increased 2.5-fold in the presence of HCO ₃ ^– . In intact perfused rat liver, HCO ₃ ^– increased both Na⁺/K⁺ pump activity and O₂ consumption. These findings indicate that, in hepatocytes, net coupled Na⁺ and HCO ₃ ^– movement is inward and represents a major determinant of Na⁺ influx and Na⁺/K⁺ pump activity. About half of hepatic Na⁺/K⁺ pump activity appears dedicated to recycling Na⁺ entering in conjunction with HCO ₃ ^– to maintain [Na⁺] _i within the physiologic range.     

Na+转运体与植物的耐盐性

简要介绍Na 转运体与植物耐盐性的研究进展。     

The Rapid-onset Dystonia Parkinsonism Mutation D923N of the Na+,K+-ATPase α3 Isoform Disrupts Na+ Interaction at the Third Na+ Site

Rapid-onset dystonia parkinsonism (RDP), a rare neurological disorder, is caused by mutation of the neuron-specific α3-isoform of Na⁺,K⁺-ATPase. Here, we present the functional consequences of RDP mutation D923N. Relative to the wild type, the mutant exhibits a remarkable ∼200-fold reduction of Na⁺ affinity for activation of phosphorylation from ATP, reflecting a defective interaction of the E₁ form with intracellular Na⁺. This is the largest effect on Na⁺ affinity reported so far for any Na⁺,K⁺-ATPase mutant. D923N also affects the interaction with extracellular Na⁺ normally driving the E₁P to E₂P conformational transition backward. However, no impairment of K⁺ binding was observed for D923N, leading to the conclusion that Asp⁹²³ is specifically associated with the third Na⁺ site that is selective toward Na⁺. The crystal structure of the Na⁺,K⁺-ATPase in E₂ form shows that Asp⁹²³ is located in the cytoplasmic half of transmembrane helix M8 inside a putative transport channel, which is lined by residues from the transmembrane helices M5, M7, M8, and M10 and capped by the C terminus, recently found involved in recognition of the third Na⁺ ion. Structural modeling of the E₁ form of Na⁺,K⁺-ATPase based on the Ca²⁺-ATPase crystal structure is consistent with the hypothesis that Asp⁹²³ contributes to a site binding the third Na⁺ ion. These results in conjunction with our previous findings with other RDP mutants suggest that a selective defect in the handling of Na⁺ may be a general feature of the RDP disorder.     

Do H+ ions obscure electrogenic Na+ and K+ binding in the E1 state of the Na,K-ATPase?

In contrast to other P-type ATPases, the Na,K-ATPase binding and release of ions on the cytoplasmic side, to the state called E1, is not electrogenic with the exception of the third Na+. Since the high-resolution structure of the closely related SR Ca-ATPase in state E1 revealed the ion-binding sites deep inside the transmembrane part of the protein, the missing electrogenicity in state E1 can be explained by an obscuring counter-movement of H+ ions. Evidence for such a mechanism is presented by analysis of pH effects on Na+ and K+ binding and by electrogenic H+ movements in the E1 conformation of the Na,K-ATPase.     

胰岛β细胞Na+通道的研究进展

Na+通道存在于胰腺胰岛β细胞上;相对其它可兴奋细胞上的Na+通道,有其特殊的生理特性和功能.研究发现β细胞上的Na+通道影响了β细胞的动作电位,并可能参与胰岛素的分泌调节.但是目前对于Na+通道在β细胞上的功能还存在着很大的争议.随着新的研究技术的发展,有必要为目前对β细胞Na+通道的研究进行综述,这对于进一步研究β细胞上Na+通道的功能具有重要的指导意义.     

How does regulatory Ca2+ regulate the Na+-Ca2+ exchanger?

Spatial and temporal regulation of intracellular Ca2+ concentrations is a fundamental requirement for life. The mammalian cardiac Na+-Ca2+ exchanger serves as the main mechanism for Ca2+ efflux after heart contraction. Exchange activity is highly regulated by intracellular Ca2+, which binds two regulatory domains (CBD1 and CBD2) and triggers the full activity of the exchanger. We solved the X-ray crystallographic structure of CBD2 in the presence and absence of Ca2+. Together with mutational analysis of the Ca2+ binding sites, this study reveals the crucial role of one of the two bound Ca2+ ions and helps propose hypotheses on the mechanism of regulation of the exchanger.     

Two types of modified cardiac Na+ channels after cytosolic interventions at the α-subunit capable of removing Na+ inactivation

Failure of inactivation is the typical response of voltage-gated Na⁺ channels to the cytosolic presence of proteolytic enzymes, protein reagents such as N-bromoacetamide (NBA) or iodate, and antibodies directed against the linker between domains III and IV of the α-subunit. The present patch clamp experiments with cardiac Na⁺ channels aimed to test the hypothesis that these interventions may provoke the occurrence of non-inactivating Na⁺ channels with distinct kinetic properties. A site-directed polyclonal antibody (anti-SLP2, target sequence 1481–1496 of the cardiac Na⁺ channel α-subunit) eliminated fast Na⁺ inactivation to induce burst activity which was accompanied by the occurrence of two open states. A deactivation process terminated channel activity during membrane depolarization proceeding with time constants of close to 40 ms (at –40 mV). NBA-modified and iodate-modified Na⁺ channels were kinetically indistinguishable from the anti-SLP2-modified type since they likewise deactivate and, thus, attain an only moderate P_o of close to 20%. This is fundamentally different from the behaviour of enzymatically-modified Na⁺ channels: after cytosolic proteolysis with α-chymotrypsin, trypsin or pronase, mean P_o during membrane depolarization amounted to approximately 40% because deactivation operated extremely slowly and less efficiently (time constants 100–200 ms at –40 mV, as a minimum) or was virtually non-operating. In-vitro cleavage of the synthetic linker sequence 1481–1496 confirmed that this part of the α-subunit provides a substrate for these peptidases or reactants for NBA but cannot be chemically modified by iodate. This iodate resistance indicates that iodate-modified Na⁺ channels are based on a structural alteration of still another region which is also involved in Na⁺ inactivation, besides the linker between domains III and IV of the α-subunit. Endogenous peptidases such as calpain did not affect Na⁺ inactivation. This stresses the stochastic nature of a kinetic peculiarity of cardiac Na⁺ channels, mode-switching to a non-inactivating mode. Received: 25 May 1996 / Accepted: 12 September 1996     

植物Na+/H+逆向转运蛋白研究进展

盐胁迫主要由Na 引起,过高的Na 浓度引起的离子毒害,渗透胁迫和K ／Na 比率的不平衡使植物新陈代谢异常,这是对大多数器官造成伤害的原因。植物抵御盐胁迫的主要方式是将细胞内过多的Na 从质膜向细胞外排放和将Na 在液泡中区隔化,这一过程是由Na ／H 逆向转运蛋白完成的。本文概述了植物中Na ／H 逆向转运蛋白的发现、特征、分子生物学方面的研究,以及Na ／H 逆向转运蛋白在植物耐盐性中的重要作用。     

黄鳝膀胱尿截流及尿Na+分析

关于黄鳝(Monopterusalbuszuiew)所谓退化性腺,作者已提供过它是膀胱的解剖学和组织学证据,并发现该器官中含有大量水样液体1,2.       

大鼠单根近端肾小管Na+-K+-ATPase活性测定方法的改进

本研究旨在对Doucet等报道的定量检测大鼠单根近端肾小管Na^＋-K^＋-ATPase活性方法进行改进。取经过Ⅱ型胶原酶消化的大鼠肾脏皮质组织,在体视显微镜下手工分离单根近端肾小管,并测量其长度,经低渗和冻融处理后与[γ-^32P]ATP共同孵育,液闪法检测从[γ-^32P]ATP解离出的^32Pi,采用修正后的公式计算Na^＋-K^＋-ATPase活性。改良法与Doucet等的方法比较,测定单根近端肾小管Na^＋-K^＋-ATPase活性无显著性差异（P〉0．05）。改进后的方法节省试剂,操作简便、省时。     

植物Na+/H+反向转运蛋白的研究进展

盐胁迫是限制植物生长发育的主要因素之一,植物Na+/H+反向转运蛋白可通过将Na+逆向转运出细胞外或将Na+区隔化于液泡中来抵制环境中过高的Na+浓度.植物中Na+/H+反向转运蛋白存在于细胞质膜和液泡膜上,现在已得到多种编码这些Na+/H+反向转运蛋白的基因,对其结构功能特性进行了大量研究,并发现将这些基因转入非抗盐植物中过量表达可提高转基因植物的抗盐性.概述了Na+/H+反向转运蛋白及其编码基因的最新研究进展.     

NaCl胁迫对阿月浑子各器官Na+、K+吸收的影响

在温室盆栽条件下,用NaCl处理新疆长果阿月浑子与美国品种Kerman,处理浓度为50、150、250和500 mmol·L-1,盐处理后5、10、20 d分别取叶、茎和根,测定各器官中Na+、K+含量及Na+/K+变化规律.研究结果表明,随着土壤盐浓度的增加,长果阿月浑子和Kerman的叶、茎和根中Na+离子含量在盐处理5、10、20 d后每个处理都表现出升高趋势;而两个品种的叶、根和茎中的K+含量在盐处理后其变化规律都不稳定.长果阿月浑子叶片、茎和根系中Na+/K+的比值在盐处理5 d和10 d都升高.Kerman的叶和根系中Na+/K+比值在处理5、10、20 d后升高;茎中Na+/K+处理10 d后升高,但20 d后没有明显变化.实验结果表明NaCl胁迫后,长果阿月浑子的叶片中Na+含量增加幅度均大于Kerman,Kerman根和茎中的Na+含量增加幅度大于长果阿月浑子;NaCl胁迫5 d和10 d后,长果阿月浑子叶片中的Na+/K+的比值高于Kerman,Kerman茎和根部中的Na+/K+的比值高于长果阿月浑子,表明了长果阿月浑子和Kerman都具有一定的耐盐性,但Kerman的耐盐性强于长果阿月浑子.由于叶片和根中Na+和Na+/K+比值的变化很稳定,可以确定为阿月浑子主要抗盐指标,茎中Na+/K+的变化规律较稳定,可以作为抗盐性参考指标.     

植物Na+/H+逆向转运蛋白研究进展

盐胁迫主要由Na+引起,过高的Na+浓度引起的离子毒害,渗透胁迫和K+/Na+比率的不平衡使植物新陈代谢异常,这是对大多数器官造成伤害的原因。植物抵御盐胁迫的主要方式是将细胞内过多的Na+从质膜向细胞外排放和将Na+在液泡中区隔化,这一过程是由Na+/H+ 逆向转运蛋白完成的。本文概述了植物中Na+/H+ 逆向转运蛋白的发现、特征、分子生物学方面的研究,以及Na+/H+ 逆向转运蛋白在植物耐盐性中的重要作用。     

Is Na + K ATPase a Myelin-Associated Enzyme?

The Na + K ATPase activity associated with purified myelin has been investigated. On the basis of marker enzyme studies, the Na + K ATPase activity of myelin was higher than could be accounted for by microsomal contamination. Fractions prepared from white matter-enriched areas of rat brain showed a threefold enrichment in Na + K ATPase activity in myelin as compared with the white matter homogenate. The ATPase activity in myelin was stimulated fourfold by treatment with sodium deoxycholate, but the activity in the whole brain homogenate and the microsomal fraction was only doubled. This discontinuity temperature for Na + K ATPase activity was significantly higher for the myelin fraction (29 degrees C) than for the microsomal fraction (21 degrees C), but the energies of activation, both above and below the discontinuity temperature, were the same for both fractions, Myelin Na + K ATPase had a lower affinity for strophanthidin than the microsomal enzyme, but both fractions were inhibited to the same extent by 10-3 M-strophanthidin. The evidence thus indicated that much of the ATPase activity of myelin is not the result of microsomal contamination. Although the possibility of axolemmal contamination cannot be ruled out conclusively, indirect evidence suggest that this is not a significant factor and that Na + K ATPase may be a myelin-associated enzyme.