NaCl胁迫对茄子嫁接苗生长和离子分布的影响

以引进日本的设施栽培专用耐盐茄子品种Torvum Vigor为砧木,栽培品种苏崎茄为接穗,通过营养液栽培对80 mmol·L-1NaCl胁迫下茄子嫁接苗和自根苗的生长和离子分布进行了比较.结果表明,(1)NaCl胁迫下嫁接苗茎伸长受抑制程度显著低于自根苗,嫁接苗根部生长旺盛;(2)嫁接苗根部贮存一定量的Na 、Cl-,地上部除叶柄外各部位Na 、Cl-含量均显著低于自根苗;(3)NaCl胁迫后嫁接苗在幼叶和根部保持较高K /Na 和Ca2 /Na 值;(4)嫁接苗根部选择吸收SK,Na、SCa,Na值和幼叶选择运输SK,Na、SCa,Na值均显著高于自根苗.因此,嫁接茄子因根部Na 、Cl-的贮存,减轻了地上部的盐离子毒害,而且K 和Ca2 在幼叶和根部的特异积累使其K /Na 值和Ca2 /Na 值提高,从而增强了耐盐性.     

四种生态型芦苇叶中离子分布对生境的生理适应

采用X射线微区分析技术 ,测定了 4种生态型芦苇 (Phragmitesaustralis (CaV .)Trin .exSteud .)叶的表皮泡状细胞、叶肉细胞和叶脉维管束鞘细胞离子的含量。结果表明 :沼泽芦苇的鞘细胞内 ,K 、Na 、Ca2 、Mg2 和Cl-分布均较叶肉细胞和泡状细胞高。沙丘芦苇的泡状细胞中Ca2 分布较叶肉细胞和鞘细胞高 ,而Mg2 在其叶肉细胞 ,以及K 、Na 和Cl- 在其鞘细胞内分布均较高。在轻度盐化草甸芦苇的叶肉细胞内分布较多的Na 和Mg2 ,而在鞘细胞内K 、Ca2 和Cl- 的分布均较叶肉细胞和泡状细胞为高。重度盐化草甸芦苇的泡状细胞内Na 和Mg2 的分布较多 ;同样 ,在叶肉细胞中K 、Ca2 和Cl- 的分布也较多。最后 ,讨论了上述各种离子在不同生态型芦苇叶内分布的状况 ,以及与其环境适应的生理意义。     

四种生态型芦苇叶中离子分布对生境的生理适应

采用X射线微区分析技术,测定了4种生态型芦苇(Phragmites australis (CaV.) Trin. exSteud.)叶的表皮泡状细胞、叶肉细胞和叶脉维管束鞘细胞离子的含量.结果表明:沼泽芦苇的鞘细胞内,K+、Na+、Ca2+、Mg2+和Cl-分布均较叶肉细胞和泡状细胞高.沙丘芦苇的泡状细胞中Ca2+分布较叶肉细胞和鞘细胞高,而Mg2+在其叶肉细胞,以及K+、Na+和Cl-在其鞘细胞内分布均较高.在轻度盐化草甸芦苇的叶肉细胞内分布较多的Na+和Mg2+,而在鞘细胞内K+、Ca2+ 和Cl-的分布均较叶肉细胞和泡状细胞为高.重度盐化草甸芦苇的泡状细胞内Na+和Mg2+的分布较多;同样,在叶肉细胞中K+、Ca2+和Cl-的分布也较多.最后,讨论了上述各种离子在不同生态型芦苇叶内分布的状况, 以及与其环境适应的生理意义.     

NaCl胁迫及钙调节对青钱柳根部组织离子分布的影响

运用X-射线微区分析法研究了NaCl胁迫及钙调节下,青钱柳〔Cyclocarya paliurus(Batal.)Iljinskaja〕幼苗根部组织中离子的浓度和分布情况。结果表明,3.0和5.0 g.L-1NaCl胁迫下,青钱柳幼苗根部各层组织中Na 和Cl-的相对含量增加,K 、Ca2 和Mg2 的相对含量降低;5.0 g.L-1NaCl胁迫下加入3.0 g.L-1Ca(NO3)2后,Na 的相对含量降低,K 的相对含量增加,Cl-的相对含量变化较小。随NaCl浓度的升高,中柱和皮层中Cl-的相对含量增高;Na 大部分沉积在皮层及表皮上;K 由皮层进入中柱的比率增大;Ca2 由表皮进入皮层的比率减少,由皮层进入中柱的比率增大。5.0 g.L-1NaCl胁迫下加入高浓度Ca(NO3)2后,Cl-的分布比率变化不明显;Na 由表皮进入皮层的比率增加,从皮层进入中柱的比率下降;K 和Ca2 的分布比率变化不明显。高浓度NaCl胁迫破坏了青钱柳幼苗根部的离子平衡,加入外源Ca(NO3)2(1.0~3.0 g.L-1)后,缓解作用不明显。     

NaCl胁迫对玉米幼苗不同器官离子含量的影响

在实验室水培条件下,研究了NaCl胁迫下玉米幼苗不同器官中Na+、K+,Ca2+,Mg 2+和Cl-含量的变化.结果表明:玉米各个部分Na+和Cl-含量、Na+/K+和Na+/Ca2+比值均随着培养液中NaCl浓度的增加而迅速提高,Na+,K+和Cl-含量的变化幅度为根系>成熟叶叶鞘>生长叶>成熟叶叶片,玉米幼苗根系最易受外界离子浓度的影响,叶片受外界环境影响较小;各器官中Ca2+、Mg2+对盐胁迫的响应不一致,NaCl胁迫使根系中Ca2+、Mg2+含量下降,成熟叶叶鞘中Mg2+含量变化规律性不明显,而NaCl胁迫下,成熟叶叶片中Ca2+、Mg2+含量增加;玉米幼苗具有拒Na+机制,具有一定的耐盐性,它的耐盐性是通过根和成熟叶叶鞘来实现的,Na+主要贮存在根系和成熟叶叶鞘中,而向成熟叶叶片和生长叶中运输较少;成熟叶叶鞘同时还具有拒Cl-能力.     

NaCl胁迫对2个杨树品系扦插苗生长及体内离子含量和运输的影响

以杨树(Populus spp.)品系“南杨1号”(Nanyang No.1)和“南杨2号”(Nanyang No.2)为实验材料,研究了不同浓度(0、75和150 mmol·L-1)NaCl胁迫条件下2个杨树品系扦插苗生长及不同器官中离子(N、P、K+、Na+、Ca2+、Mg2+和Cl-)含量与运输的差异.结果表明:随NaCl浓度的提高,2个杨树品系的单株干质量以及“南杨1号”的根冠比均逐渐降低,但“南杨2号”的根冠比呈现先增大后减小的趋势;除P和Mg2+含量外,2个品系根、新生枝条和叶片中营养元素的含量均逐渐降低,Na+和Cl-含量以及Na+/K+和Na+/Ca2+比值均逐渐增加,但Na+和Cl-含量以及Na+/K+和Na+/Ca2+比值的增幅在根中均最高、在叶片中均最小.在150 mmol·L-1NaCl胁迫条件下,2个品系的单株干质量和根冠比以及根、新生枝条和叶片中N、P、K+、Ca2+和Mg2+含量均最低,Na+和Cl-含量以及Na+/K+和Na+/Ca2+比值均最高,且与对照有显著差异.在NaCl胁迫条件下,2个品系从根到新生枝条、从新生枝条到叶片的离子运输相对选择性比率RSK+,Na+和RSCa2+,Na+基本上均小于对照,其中,从根到新生枝条的RSx+,Na+和RSCa2+,Na+均大于从新生枝条到叶片.总体上看,在NaCl胁迫条件下“南杨2号”的单株干质量和根冠比、各器官的N和P含量、不同器官间的RSK+Na+和RSCa2+,Na+均高于“南杨1号”,“南杨2号”各器官的Na+和Cl-含量以及Na+/K+和Na+/Ca2+比值的增幅均低于“南杨1号”.综合分析结果表明:NaCl胁迫对2个杨树品系扦插苗的生长及体内离子的分布及运输均有一定的影响,但总体上看,“南杨2号”对NaCl胁迫的耐性优于“南杨1号”.     

短杆菌肽通道内离子K~＋，Na~＋，Li~＋与水的相关性

基于右手螺旋短杆菌肽Ａ离子通道模型,利用分子动力学计算机模拟方法研究了通道内离子Ｋ＋,Ｎａ＋,Ｌｉ＋与水分子的相关性．     

NaCl胁迫对坪山柚、福橘实生苗矿质营养吸收特性的影响

通过沙培和水培实验,采用原子吸收分光光度计法和离子吸收动力学方法,对NaCl胁迫下坪山柚(Citrus grandis Osbeck 'Pingshanyou')和福橘(C. reticulata Blanco 'Fuju')实生苗矿质营养吸收特性进行研究. 结果表明, 随NaCl浓度增加, 坪山柚和福橘幼苗地上部及根部Na 、 Cl-含量明显增加, K 、 Ca2 含量降低. 相同浓度NaCl胁迫下, 坪山柚地上部及根部Na 、 Cl-含量分别低于福橘, 而根部K 、 Ca2 含量及地上部K 含量、 K /Na 值、 Ca2 /Na 值均高于福橘.NaCl胁迫下,坪山柚、福橘幼苗Na 吸收量高于Cl-吸收量,Na 及 Cl-的吸收速率均随NaCl浓度的增大而增加.相同浓度NaCl胁迫下,坪山柚对Na 、Cl-的吸收速率低于福橘,这与坪山柚对Na 、Cl-的吸收具有较大的Km值及较小的Vmax值一致,表明坪山柚幼苗比福橘幼苗耐盐性强.     

NaCl胁迫对营养液栽培嫁接黄瓜生物量及离子分布的影响

以‘帝王新土佐’南瓜为砧木,以‘新泰密刺’黄瓜品种为接穗,在100 mmol.L-1NaCl胁迫下,对黄瓜嫁接和自根植株生物量及各器官离子含量的差异进行了研究。结果表明,(1)NaCl胁迫下,嫁接和自根植株生物量积累均受到显著抑制,嫁接植株受抑制较轻。(2)NaCl胁迫后,黄瓜嫁接、自根植株各器官Na 、Cl-含量均显著高于对照;嫁接植株除根系Na 含量显著高于自根植株外,其余各器官均显著低于自根植株;嫁接植株老叶、叶柄和根系的Cl-含量显著低于自根植株。(3)嫁接植株的根是主要的聚Na 部位,茎是主要的聚Cl-部位。(4)嫁接植株幼叶和根的K 、Ca2 、Mg2 含量均显著高于自根植株,嫁接植株的K /Na 、Ca2 /Na 、Mg2 /Na 值也均显著高于自根植株。以上结果证明,黄瓜嫁接植株根系和茎中盐离子的含量较高且对K 、Ca2 、Mg2 的选择性吸收、运输能力较强,在器官水平上盐分离子分布的区域化优于自根植株,从而使嫁接植株耐盐性强于自根植株。     

外源EBR对NaCl胁迫下燕麦幼苗无机离子吸收、运输和分配的影响

为了探讨外源2,4-表油菜素内酯(2,4-epibrassinolide,EBR)诱导燕麦(Avena sativa L.)幼苗抗盐性的效果及其生理调节机制,以"青引2号"和"加燕2号"燕麦为材料,研究NaCl胁迫下施用外源EBR对燕麦幼苗无机离子吸收、运输和分配的影响。结果表明:100mmol·L-1NaCl胁迫下,"青引2号"和"加燕2号"燕麦幼苗叶片和根系中的Na+、Cl-含量均显著升高,对阳离子的吸收产生了拮抗作用,导致燕麦幼苗叶片和根系中的K+、Ca2+、Mg2+、Mn2+、Fe2+、Zn2+、Cu2+含量显著降低,离子稳态平衡被打破; 100 mmol·L-1NaCl胁迫下,施用0.01μmol·L-1外源EBR后,"青引2号"和"加燕2号"燕麦幼苗叶片和根系中的Na+和Cl-含量显著降低,促进了燕麦幼苗根系对K+、Ca2+、Mg2+、Fe2+、Mn2+、Cu2+和Zn2+的吸收,叶片和根系中K+/Na+、Cl-/Na+、Ca2+/Na+、Mg2+/Na+、Fe2+/Na+、Mn2+/Na+、Cu2+/Na+和Zn2+/Na+显著升高,并且有效调控燕麦幼苗体内无机离子的运输比和阳离子的运输选择性比率,离子稳态重新达到平衡状态;说明外源EBR能够缓解NaCl胁迫下Na+和Cl-对燕麦幼苗所造成的离子毒害作用,有效调控燕麦幼苗对无机离子的选择性吸收、运输和分配,对维持燕麦幼苗体内的离子稳态平衡具有正向调控作用。     

Molecular dynamics simulations of the gramicidin A-dimyristoylphosphatidylcholine system with an ion in the channel pore region

To investigate the process of ion permeation in an ion channel systematically, we performed molecular dynamics (MD) simulations on a gramicidin A (GA)-phospholipid model system with an ion in the channel pore region. Each of the three types of ions (Ca2+, Na+ Cl-) was placed at five different positions along the channel axis by replacing a water molecule. MD simulations were performed on each system at constant pressure and constant temperature. The MD trajectories showed that the Ca2+ and Na+ ions could stably fluctuate in the pore region, but the Cl- ion was pushed out because of the unfavorable interaction with the channel. This result is consistent with experimental data. It was also found that the conformation of the GA channel underwent a significant change due to the presence of the ion, and the two ends of the GA monomer were more flexible than its middle region. In particular, the dramatic change of local pore radius near the ion indicated this kind of deformation. The strong interaction between the ion and carbonyl oxygen atoms of GA was the major contributor to this change. Furthermore, it was found that the ethanolamine group of the GA molecule was the most flexible group in the GA channel and often observed to block the entrance of GA. These results imply that the deformation of channel structure plays a very important factor in ion permeation, and the ethanolamine group may play a key role in regulating ion entry into the pore. In conclusion, our results indicate that the ion has a dominant influence on the structure of the GA channel and that the flexibility of the ion channel is a crucial factor in the ion permeation process.     

Bead-like passage of chloride ions through ClC chloride channels

The ClC chloride channels control the ionic composition of the cytoplasm and the volume of cells, and regulate electrical excitability. Recently, it has been proposed that prokaryotic ClC channels are H+-Cl- exchange transporter. Although X-ray and molecular dynamics (MD) studies of bacterial ClC channels have investigated the filter open-close and ion permeation mechanism of channels, details have remained unclear. We performed MD simulations of ClC channels involving H+, Na+, K+, or H3O+ in the intracellular region to elucidate the open-close mechanism, and to clarify the role of H+ ion an H+-Cl- exchange transporter. Our simulations revealed that H+ and Na+ caused channel opening and the passage of Cl- ions. Na+ induced a bead-like string of Cl- -Na+-Cl--Na+-Cl- ions to form and permeate through ClC channels to the intracellular side with the widening of the channel pathway.     

Calcium channel selectivity for divalent and monovalent cations. Voltage and concentration dependence of single channel current in ventricular heart cells

Single channel and whole cell recordings were used to study ion permeation through Ca channels in isolated ventricular heart cells of guinea pigs. We evaluated the permeability to various divalent and monovalent cations in two ways, by measuring either unitary current amplitude or reversal potential (Erev). According to whole cell measurements of Erev, the relative permeability sequence is Ca2+ greater than Sr2+ greater than Ba2+ for divalent ions; Mg2+ is not measurably permeant. Monovalent ions follow the sequence Li+ greater than Na+ greater than K+ greater than Cs+, and are much less permeant than the divalents. These whole cell measurements were supported by single channel recordings, which showed clear outward currents through single Ca channels at strong depolarizations, similar values of Erev, and similar inflections in the current-voltage relation near Erev. Information from Erev measurements stands in contrast to estimates of open channel flux or single channel conductance, which give the sequence Na+ (85 pS) greater than Li+ (45 pS) greater than Ba2+ (20 pS) greater than Ca2+ (9 pS) near 0 mV with 110-150 mM charge carrier. Thus, ions with a higher permeability, judged by Erev, have lower ion transfer rates. In another comparison, whole cell Na currents through Ca channels are halved by less than 2 microM [Ca]o, but greater than 10 mM [Ca]o is required to produce half-maximal unitary Ca current. All of these observations seem consistent with a recent hypothesis for the mechanism of Ca channel permeation, which proposes that: ions pass through the pore in single file, interacting with multiple binding sites along the way; selectivity is largely determined by ion affinity to the binding sites rather than by exclusion by a selectivity filter; occupancy by only one Ca ion is sufficient to block the pore's high conductance for monovalent ions like Na+; rapid permeation by Ca ions depends upon double occupancy, which only becomes significant at millimolar [Ca]o, because of electrostatic repulsion or some other interaction between ions; and once double occupancy occurs, the ion-ion interaction helps promote a quick exit of Ca ions from the pore into the cell.     

A nanosecond molecular dynamics study of antiparallel d(G)7 quadruplex structures: effect of the coordinated cations

Nanosecond scale molecular dynamics simulations have been performed on antiparallel Greek key type d(G7) quadruplex structures with different coordinated ions, namely Na+ and K+ ion, water and Na+ counter ions, using the AMBER force field and Particle Mesh Ewald technique for electrostatic interactions. Antiparallel structures are stable during the simulation, with root mean square deviation values of approximately 1.5 A from the initial structures. Hydrogen bonding patterns within the G-tetrads depend on the nature of the coordinated ion, with the G-tetrad undergoing local structural variation to accommodate different cations. However, alternating syn-anti arrangement of bases along a chain as well as in a quartet is maintained through out the MD simulation. Coordinated Na+ ions, within the quadruplex cavity are quite mobile within the central channel and can even enter or exit from the quadruplex core, whereas coordinated K+ ions are quite immobile. MD studies at 400K indicate that K+ ion cannot come out from the quadruplex core without breaking the terminal G-tetrads. Smaller grooves in antiparallel structures are better binding sites for hydrated counter ions, while a string of hydrogen bonded water molecules are observed within both the small and large grooves. The hydration free energy for the K+ ion coordinated structure is more favourable than that for the Na+ ion coordinated antiparallel quadruplex structure.     

Hybrid MD-Nernst Planck model of α-hemolysin conductance properties

Motivated by experiments in which an applied electric field translocates polynucleotides through an α-hemolysin protein channel causing ionic current transient blockade, a hybrid simulation model is proposed to predict the conductance properties of the open channel. Time scales corresponding to ion permeation processes are reached using the Poisson–Nernst–Planck (PNP) electro-diffusion model in which both solvent and local ion concentrations are represented as a continuum. The diffusion coefficients of the ions (K⁺ and Cl^?) input in the PNP model are, however, calculated from all-atom molecular dynamics (MD). In the MD simulations, a reduced representation of the channel is used. The channel is solvated in a 1?M KCl solution, and an external electric field is applied. The pore specific diffusion coefficients for both ionic species are reduced 5–7 times in comparison to bulk values. Significant statistical variations (17–45%) of the pore-ions diffusivities are observed. Within the statistics, the ionic diffusivities remain invariable for a range of external applied voltages between 30 and 240?mV. In the 2D-PNP calculations, the pore stem is approximated by a smooth cylinder of radius ～9?Å with two constriction blocks where the radius is reduced to ～6?Å. The electrostatic potential includes the contribution from the atomistic charges. The MD-PNP model shows that the atomic charges are responsible for the rectifying behaviour and for the slight anion selectivity of the α-hemolysin pore. Independent of the hierarchy between the anion and cation diffusivities, the anionic contribution to the total ionic current will dominate. The predictions of the MD-PNP model are in good agreement with experimental data and give confidence in the present approach of bridging time scales by combining a microscopic and macroscopic model.     

Ions and counterions in a biological channel: a molecular dynamics simulation of OmpF porin from Escherichia coli in an explicit membrane with 1 M KCl aqueous salt solution

A 5 ns all-atom molecular dynamics trajectory of Escherichia coli OmpF porin embedded in an explicit dimyristoyl-phosphatidylcholine (DMPC) bilayer bathed by a 1 M [KCl] aqueous salt solution is generated to explore the microscopic details of the mechanism of ion permeation. The atomic model includes the OmpF trimer, 124 DMPC, 13470 water molecules as well as 231 K+ and 201 Cl-, for a total of 70,693 atoms. The structural and dynamical results are in excellent agreement with the X-ray data. The global root-mean-square deviation of the backbone atoms relative to the X-ray structure is 1.4 A. A cluster of three fully charged arginine (Arg42, Arg82, and Arg132) facing two acidic residues (Asp113 and Glu117) on L3 in the narrowest part of the aqueous pore is observed to be very stable in the crystallographic conformation. In this region of the pore, the water molecules are markedly oriented perpendicular to the channel axis due to the strong transversal electrostatic field arising from those residues. On average the size of the pore is smaller during the simulation than in the X-ray structure, undergoing small fluctuations. No large movements of loop L3 leading to a gating of the pore are observed. Remarkably, it is observed that K+ and Cl- follow two well-separated average pathways spanning over nearly 40 A along the axis of the pore. In the center of the monomer, the two screw-like pathways have a left-handed twist, undergoing a counter-clockwise rotation of 180 degrees from the extracellular vestibule to the pore periplasmic side. In the pore, the dynamical diffusion constants of the ions are reduced by about 50% relative to their value in bulk solvent. Analysis of ion solvation across the channel reveals that the contributions from the water and the protein are complementary, keeping the total solvation number of both ions nearly constant. Unsurprisingly, K+ have a higher propensity to occupy the aqueous pore than Cl-, consistent with the cation selectivity of the channel. However, further analysis suggests that ion-ion pairs play an important role. In particular, it is observed that the passage of Cl- occurs only in the presence of K+ counterions, and isolated K+ can move through the channel and permeate on their own. The presence of K+ in the pore screens the negative electrostatic potential arising from OmpF to help the translocation of Cl- by formation of ion pairs.     

Potassium and sodium binding to the outer mouth of the K+ channel.

Molecular dynamics simulations of the K+ channel from Streptomyces lividans (KcsA channel) were performed in a membrane-mimetic environment with Na+ and K+ in different initial locations. The structure of the channel remained stable and well preserved for simulations lasting up to 1.5 ns. Salt bridges between Asp80 and Arg89 of neighboring subunits, not detected in the X-ray structure, enhanced the stability of the tetrameric structure. Na+ or K+ ions located in the channel vestibule lost part of their hydration shell and diffused into the channel inner pore in less than a few hundred picoseconds. This powerful catalytic action was caused by strong electrostatic interactions with Asp80 and Glu71. The hydration state of the metal ions turned out to depend significantly on the conformational flexibility of the channel. Furthermore, Na+ entered the channel inner pore bound to more water molecules than K+. The different hydration state of the two ions may be a determinant factor in the ion selectivity of the channel.     

Conduction of Monovalent and Divalent Cations in the Slow Vacuolar Channel

The conduction properties of individual physiologically important cations Na+, K+, Mg2+, and Ca2+ were determined in the slowly activating (SV) channel of sugar beet vacuoles. Current-voltage relationships of the open channel were measured on excised tonoplast patches in a continuous manner by applying a +/-140 mV ramp-wave protocol. Applying KCl gradients of either direction across the patch we have determined that the relative Cl- to K+ permeability was < or =1%. Symmetrical increase of the concentration of tested cation caused an increase of the single channel conductance followed by saturation. Fitting of binding isotherms at zero voltage to the Michaelis-Menten equation resulted in values of maximal conductance of 300, 385, 18, and 13 pS, and of apparent dissociation constants of 64, 103, 0.04, and 0.08 mm for Na+, K+, Mg2+, and Ca2+, respectively. Deviations from the single-ion occupancy mechanism are documented, and alternative models of permeation are discussed. The magnitude of currents carried by divalent cations at low concentrations can be explained by an unrealistically wide (approximately 140 A) radius of the pore entrance. We propose instead a fixed negative charge in the pore vestibules, which concentrates the cations in their proximity. The conduction properties of the SV channel are compared with reported characteristics of voltage-dependent Ca2+-permeable channels, and consequences for a possible reduction of postulated multiplicity of Ca2+ pathways across the tonoplast are drawn.     

Ion solvation and structural stability in a sodium channel investigated by molecular dynamics calculations

The stability and ion binding properties of the homo-tetrameric pore domain of a prokaryotic, voltage-gated sodium channel are studied by extensive all-atom molecular dynamics simulations, with the channel protein being embedded in a fully hydrated lipid bilayer. It is found that Na(+) ion presents in a mostly hydrated state inside the wide pore of the selectivity filter of the sodium channel, in sharp contrast to the nearly fully dehydrated state for K(+) ions in potassium channels. Our results also indicate that Na(+) ions make contact with only one or two out of the four polypeptide chains forming the selectivity filter, and surprisingly, the selectivity filter exhibits robust stability for various initial ion configurations even in the absence of ions. These findings are quite different from those in potassium channels. Furthermore, an electric field above 0.5V/nm is suggested to be able to induce Na(+) permeation through the selectivity filter.     

Potential of mean force calculations on an L-type calcium channel model

To understand the mechanisms of Na(+)/Li(+) permeation at submicromolar Ca(2+) concentrations, Na(+)/Li(+) blocking at higher Ca(2+) concentrations (10(-6)-10(-4) M) and Ca(2+) permeation at millimolar Ca(2+) concentrations, we used our recently described L-type calcium channel model. For this purpose, we obtained potential of mean force (pmf) curves for the position change of one Na(+) and one Ca(2+) ion inside the channel and for the position change of a second Ca(2+) ion when the EEEE locus is coordinated to Ca(2+). The pmf curves suggest that (i) at submicromolar Ca(2+) concentrations, because of the low velocity of Ca(2+) entry in the channel, monovalent ion flux occurs; (ii) at Ca(2+) concentrations between 10(-6) and 10(-4) M, thermodynamic equilibrium between the channel and Ca(2+) is achieved; as the coordination of Ca(2+) with the locus is more favorable than the coordination of Na(+), the monovalent ion flux is blocked; and (iii) to put a second Ca(2+) ion inside the channel at an appropriate rate, the Ca(2+) concentration should reach millimolar levels. Nevertheless, the entry of a second Ca(2+) is thermodynamically unfavorable, indicating that the competition of two Ca(2+) ions for the locus leads to Ca(2+) permeation.