scholarly journals The permeability of the sodium channel in Myxicola to the alkali cations.

1976 ◽  
Vol 68 (3) ◽  
pp. 327-340 ◽  
Author(s):  
G A Ebert ◽  
L Goldman
Keyword(s):  
Field Equation ◽  
Reversal Potential ◽  
Structural Features ◽  
Selectivity Filter ◽  
Na Channel ◽  
Constant Field ◽  
Relative Permeabilities ◽  
Alkali Cations ◽  
Giant Axons ◽  
Channel Selectivity

Relative permeabilities to the alkali cations were determined, from the reversal potential (VRev), for the Na channel of internally perfused voltage-clamped Myxicola giant axons. PLi/PNa and PK/PNa are 0.94 and 0.076, respectively. Rb and Cs are not measurably permeant. VRev vs. the internal Na activity was well described by the constant field equation over a 300-fold range of internal Na concentrations. In agreement with findings on squid axons, the PK/PNa was found to increase when the K content of the internal perfusate was reduced (equivalent per equivalent substitution with TMA). Internal Rb and Cs also decreased the PK/PNa. The order of effectiveness of internal K, Rb, and Cs in increasing the Na selectivity of the Na channel was Cs greater than Rb greater than or equal to K. External Li increases the PK/PNa but this may be due to the formation of LiF internally. It may be that substances do not have to traverse the channel in order to affect the selectivity filter. Evidence is presented which suggests that the selectivity of the Na channel may be higher for Na in intact as compared to perfused giant axons. It was concluded that the channel selectivity properities do not reflect only some fixed structural features of the channel, but the selectivity filter has a labile organization.

scholarly journals Effect of acetylcholine on postjunctional membrane permeability in eel electroplaque.

1977 ◽  
Vol 70 (1) ◽  
pp. 23-36 ◽  
Author(s):  
N L Lassignal ◽  
A R Martin
Keyword(s):  
Field Equation ◽  
Membrane Potential ◽  
Membrane Permeability ◽  
Ionic Composition ◽  
Reversal Potential ◽  
Resting Potential ◽  
Constant Field ◽  
Free Solution ◽  
Positive Direction ◽  
External K

Acetylcholine (ACh) was applied iontophoretically to the innervated face of isolated eel electroplaques while the membrane potential was being recorded intracellularly. At the resting potential (about -85 mV) application of the drug produced depolarizations (ACh potentials) of 20 mV or more which became smaller when the membrane was depolarized and reversed in polarity at about zero membrane potential. The reversal potential shifted in the negative direction when external Na+ was partially replaced by glucosamine. Increasing external K+ caused a shift of reversal potential in the positive direction. It was concluded that ACh increased the permeability of the postjunctional membrane to both ions. Replacement of Cl- by propionate had no effect on the reversal potential. In Na+-free solution containing glucosamine the reversal potential was positive to the resting potential, suggesting that ACh increased the permeability to glucosamine. Addition of Ca++ resulted in a still more positive reversal potential, indicating an increased permeability to Ca++ as well. Analysis of the results indicated that the increases in permeability of the postjunctional membrane to K+, Na+, Ca++, and glucosamine were in the ratios of approximately 1.0:0.9:0.7:0.2, respectively. With these permeability ratios, all of the observed shifts in reversal potential with changes in external ionic composition were predicted accurately by the constant field equation.

scholarly journals Leak Current Rectification in Myxicola Giant Axons

1969 ◽  
Vol 54 (6) ◽  
pp. 755-764 ◽  
Author(s):  
L. Goldman ◽  
L. Binstock
Keyword(s):  
Field Equation ◽  
Membrane Potential ◽  
Resting Membrane Potential ◽  
Constant Field ◽  
Leak Current ◽  
Giant Axons ◽  
Time To Peak ◽  
Zero Current ◽  
Current Voltage ◽  
Current Voltage Curve

Early leak current, i.e. for times similar to the time to peak of the transient current was measured in Myxicola giant axons in the presence of tetrodotoxin. The leak current-voltage relation rectifies, showing more current for strong depolarizing pulses than expected from symmetry around the holding potential. A satisfactory practical approximation for most leak corrections is constant resting conductance. The leak current-voltage curve rectifies less than expected from the constant field equation. These curves cannot be reconstructed by summing the constant field currents for sodium and potassium using a PNa/PK ratio obtained in the usual way, from zero current constant field fits to resting membrane potential data. Nor can they be reconstructed by summing the constant field current for potassium with that for any other single ion. They can be reconstructed, however, by summing the constant field current for potassium with a constant conductance component. It is concluded that the leak current and the resting membrane potential, therefore, are determined by multiple ionic components, at least three and possibly many. Arguments are presented suggesting that ion permeability ratios obtained in the usual way, by fitting the constant field equation to resting membrane potential data should be viewed with skepticism.

scholarly journals Ionic selectivity of the sodium channel of frog skeletal muscle.

1976 ◽  
Vol 67 (3) ◽  
pp. 295-307 ◽  
Author(s):  
D T Campbell
Keyword(s):  
Node Of Ranvier ◽  
Selectivity Filter ◽  
Na Channel ◽  
Ion Permeability ◽  
Ionic Selectivity ◽  
Semitendinosus Muscle ◽  
Channel Selectivity ◽  
Inward Currents ◽  
Channel Currents ◽  
Reversal Potentials

The ionic selectivity of the Na channel to a variety of metal and organic cations is studied in frog semitendinosus muscle. Na channel currents are measured under voltage clamp conditions in fibers bathed in solutions with all Na+ replaced by a test ion. Permeability ratios are calculated from measured reversal potentials using the Goldman-Hodgkin-Katz equation. The permeability sequence was Na+ approximately Li+ approximately hydroxylammonium greater than hydrazinium greater than ammonium greater than guanidinium greater than K+ greater than aminoguanidinium in the ratios 1:0.96:0.94:0.31:0.11:0.093:0.048:0.031. No inward currents were observed for Ca++, methylammonium, methylguanidinium, tetraethylammonium, and tetramethylammonium. The results are consistent with the Hille model of the Na channel selectivity filter of the node of Ranvier and suggest that the selectivity filter of the two channels is the same.

scholarly journals Charged Residues between the Selectivity Filter and S6 Segments Contribute to the Permeation Phenotype of the Sodium Channel

1999 ◽  
Vol 115 (1) ◽  
pp. 81-92 ◽  
Author(s):  
Ronald A. Li ◽  
Patricio Vélez ◽  
Nipavan Chiamvimonvat ◽  
Gordon F. Tomaselli ◽  
Eduardo Marbán
Keyword(s):  
Single Channel ◽  
Structural Features ◽  
Selectivity Filter ◽  
Na Channel ◽  
Ion Permeation ◽  
Structural Topology ◽  
Functional Roles ◽  
Charged Residues ◽  
Domain Iii ◽  
And Function

The deep regions of the Na+ channel pore around the selectivity filter have been studied extensively; however, little is known about the adjacent linkers between the P loops and S6. The presence of conserved charged residues, including five in a row in domain III (D-III), hints that these linkers may play a role in permeation. To characterize the structural topology and function of these linkers, we neutralized the charged residues (from position 411 in D-I and its homologues in D-II, -III, and -IV to the putative start sites of S6) individually by cysteine substitution. Several cysteine mutants displayed enhanced sensitivities to Cd2+ block relative to wild-type and/or were modifiable by external sulfhydryl-specific methanethiosulfonate reagents when expressed in TSA-201 cells, indicating that these amino acids reside in the permeation pathway. While neutralization of positive charges did not alter single-channel conductance, negative charge neutralizations generally reduced conductance, suggesting that such charges facilitate ion permeation. The electrical distances for Cd2+ binding to these residues reveal a secondary “dip” into the membrane field of the linkers in domains II and IV. Our findings demonstrate significant functional roles and surprising structural features of these previously unexplored external charged residues.

scholarly journals Sodium channel selectivity and conduction: Prokaryotes have devised their own molecular strategy

2014 ◽  
Vol 143 (2) ◽  
pp. 157-171 ◽  
Author(s):  
Rocio K. Finol-Urdaneta ◽  
Yibo Wang ◽  
Ahmed Al-Sabi ◽  
Chunfeng Zhao ◽  
Sergei Y. Noskov ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Tight Binding ◽  
Selectivity Filter ◽  
Alkali Cations ◽  
Molecular Feature ◽  
Ion Interactions ◽  
Channel Selectivity ◽  
Potentials Of Mean Force ◽  
In The Wild ◽  
Nav Channels

Striking structural differences between voltage-gated sodium (Nav) channels from prokaryotes (homotetramers) and eukaryotes (asymmetric, four-domain proteins) suggest the likelihood of different molecular mechanisms for common functions. For these two channel families, our data show similar selectivity sequences among alkali cations (relative permeability, Pion/PNa) and asymmetric, bi-ionic reversal potentials when the Na/K gradient is reversed. We performed coordinated experimental and computational studies, respectively, on the prokaryotic Nav channels NaChBac and NavAb. NaChBac shows an “anomalous,” nonmonotonic mole-fraction dependence in the presence of certain sodium–potassium mixtures; to our knowledge, no comparable observation has been reported for eukaryotic Nav channels. NaChBac’s preferential selectivity for sodium is reduced either by partial titration of its highly charged selectivity filter, when extracellular pH is lowered from 7.4 to 5.8, or by perturbation—likely steric—associated with a nominally electro-neutral substitution in the selectivity filter (E191D). Although no single molecular feature or energetic parameter appears to dominate, our atomistic simulations, based on the published NavAb crystal structure, revealed factors that may contribute to the normally observed selectivity for Na over K. These include: (a) a thermodynamic penalty to exchange one K+ for one Na+ in the wild-type (WT) channel, increasing the relative likelihood of Na+ occupying the binding site; (b) a small tendency toward weaker ion binding to the selectivity filter in Na–K mixtures, consistent with the higher conductance observed with both sodium and potassium present; and (c) integrated 1-D potentials of mean force for sodium or potassium movement that show less separation for the less selective E/D mutant than for WT. Overall, tight binding of a single favored ion to the selectivity filter, together with crucial inter-ion interactions within the pore, suggests that prokaryotic Nav channels use a selective strategy more akin to those of eukaryotic calcium and potassium channels than that of eukaryotic Nav channels.

scholarly journals Slowing of sodium current inactivation by ruthenium red in snail neurons.

1982 ◽  
Vol 80 (4) ◽  
pp. 485-497 ◽  
Author(s):  
J R Stimers ◽  
L Byerly
Keyword(s):  
Sodium Current ◽  
Reversal Potential ◽  
Membrane Resistance ◽  
Ruthenium Red ◽  
Na Channel ◽  
Constant Field ◽  
Na Current ◽  
High Concentration ◽  
Voltage Dependent ◽  
Current Flows

The effects of ruthenium red (RuR) were tested on the membrane currents of internally perfused, voltage-clamped nerve cell bodies from the snail Limnea stagnalis. Bath application of nanomolar concentrations of RuR produces a prolonged Na current that decays approximately 40 times slower than the normal Na current in these cells. The relationship between the reversal potential for the prolonged Na current and the intracellular concentration of Na+ agrees well with the constant-field equation, assuming a small permeability for Cs+. Because a strong correlation was found between the magnitude of the normal Na current and that of the prolonged Na current, it is concluded that the prolonged Na current flows through the normal Na channels. This conclusion is supported by the similar selectivities, voltage dependencies, and tetrodotoxin (TTX) sensitivities of these two currents. This action of RuR to slow the inactivation of the Na channel was not observed at concentrations below 1 nM, but was complete at 10 nM. When the concentration of RuR is increased to 0.1 mM, the Ca current in these cells is blocked; but at this high concentration RuR also reduces the outward voltage-dependent currents and resting membrane resistance. Therefore, RuR is not a good Ca blocker because of its lack of specificity. However, its action of slowing Na current inactivation is very specific and could prove to be useful in studying the inactivation of the Na channel.

scholarly journals Instantaneous ion configurations in the K+ion channel selectivity filter revealed by 2D IR spectroscopy

Science ◽  
2016 ◽  
Vol 353 (6303) ◽  
pp. 1040-1044 ◽  
Author(s):  
Huong T. Kratochvil ◽  
Joshua K. Carr ◽  
Kimberly Matulef ◽  
Alvin W. Annen ◽  
Hui Li ◽  
...  
Keyword(s):  
Ir Spectroscopy ◽  
Ion Channel ◽  
Selectivity Filter ◽  
2D Ir ◽  
Channel Selectivity ◽  
Ion Channel Selectivity

An Investigation of the Electrogenic Sodium Pump in Snail Neurones, Using the Constant-Field Theory

1969 ◽  
Vol 51 (1) ◽  
pp. 181-201
Author(s):  
R. B. MORETON
Keyword(s):  
Field Equation ◽  
Sodium Pump ◽  
Membrane Resistance ◽  
Resting Potential ◽  
Constant Field ◽  
Sodium Ions ◽  
Intracellular Potassium ◽  
Electrogenic Sodium Pump ◽  
Snail Neurones ◽  
Intracellular Potassium Concentration

1. Sodium ions injected into giant neurones of Helix aspersa by diffusion from low-resistance microelectrodes caused hyperpolarization of the cells. Under these conditions the behaviour of the resting potential could be described by a modified ‘constant-field’ equation, including a term representing the effect of a potassiumsensitive, electrogenic sodium pump. 2. Exposure to potassium-free solution, ouabain or cyanide abolished the hyperpolarization, and caused a gradual fall in the intracellular potassium concentration, as estimated from the constant-field equation. 3. Assuming that this fall was due to replacement of intracellular potassium by injected sodium ions, it was possible to calculate the rates of injection and pumping of sodium ions, and, using the measured membrane resistance of the cell, the hyperpolarization which the sodium pump could cause, if it were electrogenic. 4. This was related to the observed hyperpolarization, supporting the view that the latter was caused by stimulation of the electrogenic sodium pump.

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