Multiple conductance states of the purified calcium release channel complex from skeletal sarcoplasmic reticulum.
The CHAPS-solubilized and purified 30S ryanodine receptor protein complex from skeletal sarcoplasmic reticulum (SR) was incorporated into planar lipid bilayers. The resulting electrical activity displayed similar responses to agents such as Ca2+, ATP, ryanodine, or caffeine as the native Ca2+ release channel, confirming the identification of the 30S complex as the Ca2+ release channel. The purified channel was permeable to monovalent ions such as Na+, with the permeability ratio PCa/PNa approximately 5, and was highly selective for cations over anions. The purified channel also showed at least four distinct conductance levels for both Na+ and Ca2+ conducting ions, with the major subconducting level in NaCl buffers possessing half the conductance value of the main conductance state. These levels may be produced by intrinsic subconductances present within the channel oligomer. Several of these conductances may be cooperatively coupled to produce the characteristic 100 +/- 10 pS unitary Ca2+ conductance of the native channel.
Liu
Q Y
QY
Department of Biochemistry, University of North Carolina School of Medicine, Chapel Hill 27599-7260.
Lai
F A
FA
Rousseau
E
E
Jones
R V
RV
Meissner
G
G
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D015220:Calcium Channels / Q000187:drug effects / Q000502:physiology*; D004533:Egtazic Acid / Q000494:pharmacology; D004553:Electric Conductivity; D007700:Kinetics; D008051:Lipid Bilayers; D009132:Muscles / Q000502:physiology*; D011950:Receptors, Cholinergic / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism* / Q000502:physiology*; D019837:Ryanodine Receptor Calcium Release Channel; D012519:Sarcoplasmic Reticulum / Q000502:physiology*
D015220:Calcium Channels; D008051:Lipid Bilayers; D011950:Receptors, Cholinergic; D019837:Ryanodine Receptor Calcium Release Channel; D004533:Egtazic Acid; D002118:Calcium