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PMID: 2429253 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Solubilisation and reconstitution of the rabbit skeletal muscle sarcoplasmic reticulum K+ channel into liposomes suitable for patch clamp studies.

Pflugers Archiv : European journal of physiology ·Vol. 407 ·No. 3 ·1986-09-00 ·Pages 341-7

Tomlins B, Williams AJ

Abstract

Sarcoplasmic reticulum (SR) membrane vesicles have been prepared from rabbit skeletal muscle and solubilised using K+ cholate. Solubilised membrane proteins were reconstituted into small asolectin liposomes by dialysis against cholate-free solution. Large liposomes were produced by freezing and thawing at -80 degrees C and room temperature, respectively. The liposomes were assayed for the SR K+ channel using the patch clamp technique. Channel density was modulated by varying protein:lipid ratios during reconstitution. Channels inserted into the membrane with a preferred orientation. The solubilised and reconstituted channel behaves ohmically over the holding potential range +/- 70 mV and has a conductance of 178.4 +/- 4.4 pS (mean +/- SE, n = 37) in 200 mM KCl. The channel has a selectivity sequence of K+ greater than NH4+ greater than Rb+ greater than Na+ and K+ conductance is blocked by hexamethonium and decamethonium. The opening probability of the reconstituted channel is voltage dependent. The conductance and gating characteristics displayed by the solubilised and reconstituted channel correlate well with those previously observed following the fusion of native SR membrane vesicles with planar phospholipid bilayers.

MeSH Terms
Animals Electrophysiology Histological Techniques Ion Channels/physiology Liposomes/classification,physiology Muscles/metabolism Potassium/metabolism Rabbits Sarcoplasmic Reticulum/metabolism Solubility
Chemicals
Ion Channels Liposomes Potassium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Tomlins B
Williams A J
References (30)
30 references, click to expand
  1. Permeability of reconstituted sarcoplasmic reticulum vesicles. Reconstitution of the K+, Na+ channel.
    Biochim Biophys Acta. 1981 Jan 22;640(2):409-18 PMID: 6260253
  2. Purification of the tetrodotoxin-binding component associated with the voltage-sensitive sodium channel from Electrophorus electricus electroplax membranes.
    Proc Natl Acad Sci U S A. 1978 Jun;75(6):2606-10 PMID: 275831
  3. The characterization of a monovalent cation-selective channel of mammalian cardiac muscle sarcoplasmic reticulum.
    J Membr Biol. 1984;80(2):191-9 PMID: 6148425
  4. Ionic selectivity, saturation, and block in a K+-selective channel from sarcoplasmic reticulum.
    J Gen Physiol. 1980 Oct;76(4):425-46 PMID: 6255062
  5. Isolated-patch recording from liposomes containing functionally reconstituted chloride channels from Torpedo electroplax.
    Proc Natl Acad Sci U S A. 1982 Dec;79(24):7749-53 PMID: 6296849
  6. Electrically gated ionic channels in lipid bilayers.
    Q Rev Biophys. 1977 Feb;10(1):1-34 PMID: 327501
  7. Decamethonium and hexamethonium block K+ channels of sarcoplasmic reticulum.
    Nature. 1980 Dec 4;288(5790):495-7 PMID: 6255336
  8. Channel-mediated monovalent cation fluxes in isolated sarcoplasmic reticulum vesicles.
    J Gen Physiol. 1984 Jun;83(6):819-39 PMID: 6330279
  9. Ion channels in liposomes.
    Annu Rev Physiol. 1984;46:549-58 PMID: 6324663
  10. Assay of proteins in the presence of interfering materials.
    Anal Biochem. 1976 Jan;70(1):241-50 PMID: 1259145
  11. Thermodynamic and kinetic studies of the gating behavior of a K+-selective channel from the sarcoplasmic reticulum membrane.
    J Gen Physiol. 1980 Oct;76(4):397-24 PMID: 6255061
  12. Conductance and selectivity properties of a substate of the rabbit sarcoplasmic reticulum channel.
    Biophys J. 1985 Apr;47(4):573-6 PMID: 2580571
  13. Isolation of the cholinergic receptor protein of Torpedo electric tissue.
    Nature. 1971 Feb 19;229(5286):554-7 PMID: 4925349
  14. Functional reconstitution of the purified sodium channel protein from rat sarcolemma.
    Proc Natl Acad Sci U S A. 1982 Jun;79(11):3651-5 PMID: 6285356
  15. Permeability of canine cardiac sarcoplasmic reticulum vesicles to K+, Na+, H+, and Cl-.
    J Biol Chem. 1982 Jul 10;257(13):7704-11 PMID: 7085644
  16. Voltage-gated cation conductance channel from fragmented sarcoplasmic reticulum: steady-state electrical properties.
    J Membr Biol. 1978 Apr 20;40(1):1-23 PMID: 650672
  17. Evidence for a K+, Na+ permeable channel in sarcoplasmic reticulum.
    J Membr Biol. 1978 Dec 15;44(2):159-86 PMID: 731686
  18. The sodium channel from rat brain. Purification and subunit composition.
    J Biol Chem. 1984 Feb 10;259(3):1667-75 PMID: 6319405
  19. Incorporation of acetylcholine receptors into liposomes. Vesicle structure and acetylcholine receptor function.
    J Biol Chem. 1982 Jun 25;257(12):7122-34 PMID: 7085620
  20. Different channel properties of Torpedo acetylcholine receptor monomers and dimers reconstituted in planar membranes.
    Proc Natl Acad Sci U S A. 1984 Oct;81(19):6222-6 PMID: 6091143
  21. Integral membrane channels: studies in model membranes.
    Physiol Rev. 1983 Oct;63(4):1209-42 PMID: 6318238
  22. Single-channel properties of the reconstituted voltage-regulated Na channel isolated from the electroplax of Electrophorus electricus.
    Proc Natl Acad Sci U S A. 1984 Sep;81(17):5594-8 PMID: 6089214
  23. Properties of proteoliposomes reconstituted with acetylcholine receptor from Torpedo californica.
    J Biol Chem. 1982 Aug 25;257(16):9372-8 PMID: 6286618
  24. Bis-quaternary ammonium blockers as structural probes of the sarcoplasmic reticulum K+ channel.
    J Gen Physiol. 1982 May;79(5):869-91 PMID: 6284862
  25. Effects of phospholipid surface charge on ion conduction in the K+ channel of sarcoplasmic reticulum.
    Biophys J. 1984 Jan;45(1):279-87 PMID: 6324908
  26. Patch-recorded single-channel currents of the purified and reconstituted Torpedo acetylcholine receptor.
    Proc Natl Acad Sci U S A. 1983 Aug;80(16):5129-33 PMID: 6308673
  27. Acetylcholine receptor in planar lipid bilayers. Characterization of the channel properties of the purified nicotinic acetylcholine receptor from Torpedo californica reconstituted in planar lipid bilayers.
    J Gen Physiol. 1984 Apr;83(4):473-96 PMID: 6144720
  28. Purification from rat sarcolemma of the saxitoxin-binding component of the excitable membrane sodium channel.
    Proc Natl Acad Sci U S A. 1980 Mar;77(3):1306-10 PMID: 6246486
  29. Reconstitution of purified acetylcholine receptors with functional ion channels in planar lipid bilayers.
    Proc Natl Acad Sci U S A. 1980 May;77(5):3057-61 PMID: 6930685
  30. Reconstitution and purification of the D-glucose transporter from human erythrocytes.
    J Biol Chem. 1977 Oct 25;252(20):7384-90 PMID: 903365
Article Info
Journal
Pflugers Archiv : European journal of physiology
Abbr.
Pflugers Arch
ISSN
0031-6768
DOI
10.1007/BF00585312
Published
1986-09-00
Pages
341-7
Language
English
Region
Germany
NLM ID
0154720
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00585312 View journal details
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