Regulation of transient Na<sup>+</sup> conductance by intra‐ and extracellular K<sup>+</sup> in the human delayed rectifier K<sup>+</sup> channel Kv1.5
Bibliographic record
Abstract
1. Significant Na+ conductance has been described in only a few native and cloned K+ channels, but has been used to characterize inactivation and K+ binding within the permeation pathway, and to refine models of K+ flux through multi-ion pores. Here we use Na+ permeation of the delayed rectifier K+ channel Kv1.5 to study extra- and intracellular K+ (K+o and K+i, respectively) regulation of conductance and inactivation, using whole-cell recording from human embryonic kidney (HEK)-293 cells. 2. Kv1.5 Na+ currents in the absence of K+o and K+i were confirmed by: (i) resistance of outward Na+ currents to dialysis by K+-free solutions; (ii) tail current reversal potential changes with Na+o with a slope of 55.8 mV per decade; (iii) block by 4-aminopyridine (50 % at 50 microM), and resistance to Cl- channel inhibition. 3. Na+ currents were transient followed by a small sustained current. An envelope test confirmed that activated Kv1.5 channels conducted Na+, and that rapid current decay reflected C-type inactivation. Sustained currents ( approximately 13 % of peak) represented Na+ flux through inactivated Kv1.5 channels. 4. K+o could modulate the maximum available Na+ conductance in the stable cell line while channels were closed. Before the first pulse of a train, increasing K+o concentration increased the subsequent Na+ conductance from approximately 15 (0 mM K+o) to 30 nS (5 mM K+o), with a Kd of 23 microM. Repeated low rate depolarizations in Na+i/Na+o solutions induced a use-dependent loss of Kv1.5 channel Na+ conductance, distinct from that caused by C-type inactivation. K+o binding that sensed little of the electric field could prevent this secondary loss of available Kv1.5 channels with a Kd of 230 microM. These two effects on conductance were both voltage independent, and had no effect on channel inactivation rate. 5. K+o concentrations >= 0.3 mM slowed the inactivation rate in a strongly voltage-dependent manner. This suggested it could compete for binding at a K+ site or sites deeper in the pore, as well as restoring the Na+ conductance. K+i was able to modulate the inactivation rate but was unable to affect conductance. 6. Mutation of arginine 487 in the outer pore region of the channel to valine (R487V) greatly reduced C-type inactivation in Na+ solutions, caused loss of channel use dependence, and prevented any conductance increase upon the addition of 0.1 mM K+o. Our results confirm the existence of a high affinity binding site at the selectivity filter that regulates inactivation, and also reveals the presence of at least one additional high affinity outer mouth site that predominantly regulates conductance of resting channels, and protects channels activated by depolarization when they conduct Na+.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".