Variability of channel subconductance states of the cardiac sodium channel induced by protease.
Conductance and subconductance levels of voltage-activated sodium channels recorded using patch clamp techniques from isolated cardiac myocytes were accurately determined using signal processing techniques. From the tabulated amplitude distributions of the conductance levels, we inferred the most likely underlying distribution by applying the method of the kernel density estimate. When myocytes were prepared by dissociation of the heart with a solution containing collagenase as the only digestive enzyme, the fully open conductance level of the channel was 23 pS, with two prominent sublevels at 8 and 16 pS (280 mM sodium). When cells were dissociated in an identical manner but with solution containing added protease, the most frequent open levels of the channel were 9 and 15 pS. In these latter recordings, the channel also opened to 22 pS, but did so only rarely. The main conductance levels in cells dissociated with protease were essentially the same as the subconductance states in cells dissociated without protease. We infer that the conductance sublevels normally seen are, within experimental errors, 1/3 and 2/3 of the fully open level, and that the proteolytic enzyme modifies the channel such that it tends to open predominantly to the subconductance levels.
Department of Physiology, University of Adelaide, Australia. dsaint@physiol.adelaide.edu.au
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D000818:Animals; D004553:Electric Conductivity; D010450:Endopeptidases / Q000494:pharmacology*; D015640:Ion Channel Gating / Q000187:drug effects*; D017136:Ion Transport / Q000187:drug effects; D016013:Likelihood Functions; D008297:Male; D008390:Markov Chains; D009124:Muscle Proteins / Q000187:drug effects* / Q000502:physiology; D009206:Myocardium / Q000737:chemistry*; D018408:Patch-Clamp Techniques; D051381:Rats; D017208:Rats, Wistar; D012964:Sodium / Q000378:metabolism*; D015222:Sodium Channels / Q000187:drug effects* / Q000502:physiology
D009124:Muscle Proteins; D015222:Sodium Channels; D012964:Sodium; D010450:Endopeptidases