Recombinant human voltage-gated skeletal muscle sodium channels are pharmacologically functional in planar lipid bilayers

2007 ◽  
Vol 22 (6) ◽  
pp. 1006-1012 ◽  
Author(s):  
Yan Li Zhang ◽  
James Dunlop ◽  
Julie E. Dalziel
Keyword(s):  
Skeletal Muscle ◽  
Lipid Bilayers ◽  
Sodium Channels ◽  
Planar Lipid Bilayers ◽  
Voltage Gated

Modulation of skeletal muscle Ca2(+)-release channel activity by sphingosine

1997 ◽  
Vol 272 (5) ◽  
pp. C1465-C1474 ◽  
Author(s):  
D. H. Needleman ◽  
B. Aghdasi ◽  
A. B. Seryshev ◽  
G. J. Schroepfer ◽  
S. L. Hamilton
Keyword(s):  
Skeletal Muscle ◽  
Lipid Bilayers ◽  
Binding Protein ◽  
Planar Lipid Bilayers ◽  
Dependent Manner ◽  
Carboxy Terminus ◽  
Release Channel ◽  
Proteolytic Fragment ◽  
High Affinity ◽  
High Affinity Site

The effect of D-erythro-C18-sphingosine (sphingosine) and related compounds on the Ca(2+)-release channel (ryanodine binding protein) was examined on rabbit skeletal muscle membranes, on the purified ryanodine binding protein, and on the channel reconstituted into planar lipid bilayers. Sphingosine inhibited [3H]ryanodine binding to sarcoplasmic reticulum (SR) membranes in a dose-dependent manner similar to published results (R. A. Sabbadini, R. Betto, A. Teresi, G. Fachechi-Cassano, and G. Salviati. J. Biol. Chem. 267: 15475-15484, 1992). The sphingolipid also inhibited [3H]ryanodine binding to the purified ryanodine binding protein. Our results demonstrate that the inhibition of [3H]ryanodine binding by sphingosine is due to an increased rate of dissociation of bound [3H]ryanodine from SR membranes and a decreased rate of association of [3H]ryanodine to the high-affinity site. Unlike other modulators of the Ca(2+)-release channel, sphingosine can remove bound [3H]ryanodine from the high-affinity site within minutes. Sphingosine increased the rate of dissociation of [3H]ryanodine bound to a solubilized proteolytic fragment derived from the carboxy terminus of the ryanodine binding protein (cleavage at Arg4475). Sphingosine also inhibited the activity of the Ca(2+)-release channel incorporated into planar lipid bilayers. Taken together, the data provide evidence for a direct effect of sphingosine on the Ca(2+)-release channel. Sphingosine is a noncompetitive inhibitor at the high-affinity ryanodine binding site, and it interacts with a site between Arg4475 and the carboxy terminus of the Ca(2+)-release channel.

Neuraminidase treatment modifies the function of electroplax sodium channels in planar lipid bilayers

Neuron ◽  
1990 ◽  
Vol 5 (5) ◽  
pp. 675-684 ◽  
Author(s):  
Esperanza Recio-Pinto ◽  
William B. Thornhill ◽  
Daniel S. Duch ◽  
Simon R. Levinson ◽  
Bernd W. Urban
Keyword(s):  
Lipid Bilayers ◽  
Sodium Channels ◽  
Planar Lipid Bilayers ◽  
Neuraminidase Treatment

scholarly journals Potassium blocks barium permeation through a calcium-activated potassium channel.

1988 ◽  
Vol 92 (5) ◽  
pp. 549-567 ◽  
Author(s):  
J Neyton ◽  
C Miller
Keyword(s):  
Skeletal Muscle ◽  
Lipid Bilayers ◽  
Dissociation Rate ◽  
Planar Lipid Bilayers ◽  
Rat Skeletal Muscle ◽  
K Channels ◽  
Calcium Activated Potassium Channel ◽  
Micromolar Range ◽  
High Conductance ◽  
External K

Single high-conductance Ca2+-activated K+ channels from rat skeletal muscle were inserted into planar lipid bilayers, and discrete blocking by the Ba2+ ion was studied. Specifically, the ability of external K+ to reduce the Ba2+ dissociation rate was investigated. In the presence of 150 mM internal K+, 1-5 microM internal Ba2+, and 150 mM external Na+, Ba2+ dissociation is rapid (5 s-1) in external solutions that are kept rigorously K+ free. The addition of external K+ in the low millimolar range reduces the Ba2+ off-rate 20-fold. Other permeant ions, such as Tl+, Rb+, and NH4+ show a similar effect. The half-inhibition constants rise in the order: Tl+ (0.08 mM) less than Rb+ (0.1 mM) less than K+ (0.3 mM) less than Cs+ (0.5 mM) less than NH4+ (3 mM). When external Na+ is replaced by 150 mM N-methyl glucamine, the Ba2+ off-rate is even higher, 20 s-1. External K+ and other permeant ions reduce this rate by approximately 100-fold in the micromolar range of concentrations. Na+ also reduces the Ba2+ off-rate, but at much higher concentrations. The half-inhibition concentrations rise in the order: Rb+ (4 microM) less than K+ (19 microM) much less than Na+ (27 mM) less than Li+ (greater than 50 mM). The results require that the conduction pore of this channel contains at least three sites that may all be occupied simultaneously by conducting ions.

scholarly journals Unitary Ca2+ Current through Mammalian Cardiac and Amphibian Skeletal Muscle Ryanodine Receptor Channels under Near-physiological Ionic Conditions

2003 ◽  
Vol 122 (4) ◽  
pp. 407-417 ◽  
Author(s):  
Claudia Kettlun ◽  
Adom González ◽  
Eduardo Ríos ◽  
Michael Fill
Keyword(s):  
Skeletal Muscle ◽  
Charge Carrier ◽  
Lipid Bilayers ◽  
Ryanodine Receptor ◽  
Skeletal Muscles ◽  
Planar Lipid Bilayers ◽  
High Concentrations

Ryanodine receptor (RyR) channels from mammalian cardiac and amphibian skeletal muscle were incorporated into planar lipid bilayers. Unitary Ca2+ currents in the SR lumen-to-cytosol direction were recorded at 0 mV in the presence of caffeine (to minimize gating fluctuations). Currents measured with 20 mM lumenal Ca2+ as exclusive charge carrier were 4.00 and 4.07 pA, respectively, and not significantly different. Currents recorded at 1–30 mM lumenal Ca2+ concentrations were attenuated by physiological [K+] (150 mM) and [Mg2+] (1 mM), in the same proportion (∼55%) in mammalian and amphibian channels. Two amplitudes, differing by ∼35%, were found in amphibian channel studies, probably corresponding to α and β RyR isoforms. In physiological [Mg2+], [K+], and lumenal [Ca2+] (1 mM), the Ca2+ current was just less than 0.5 pA. Comparison of this value with the Ca2+ flux underlying Ca2+ sparks suggests that sparks in mammalian cardiac and amphibian skeletal muscles are generated by opening of multiple RyR channels. Further, symmetric high concentrations of Mg2+ substantially reduced the current carried by 10 mM Ca2+ (∼40% at 10 mM Mg2+), suggesting that high Mg2+ may make sparks smaller by both inhibiting RyR gating and reducing unitary current.

Sign in / Sign up

Export Citation Format

Share Document