Structural Studies of Human Plasma Membrane Anion Exchanger, AE1, Expressed in <i>Saccharomyces cerevisiae</i>
Bibliographic record
Abstract
Anion exchanger 1 (AE1), which is expressed in the renal and erythrocyte membranes, functions to exchange chloride for bicarbonate across the plasma membrane. While there is a high‐resolution structure of the N‐terminal cytoplasmic domain of AE1, there is no high‐resolution structure of the 55 kDa AE1 membrane domain. We have developed an expression and purification system for AE1, which takes advantage of S. cerevisiae as a large‐scale expression system of properly folded homogeneous AE1. The expression construct (AE1‐Rho) consists of the AE1 membrane domain, amino acids 388 to 911 of human AE1, followed by the 9 C‐terminal amino acids of rhodopsin. Milligram quantities of AE1‐Rho have been purified in the presence of the detergent fos‐choline, by a 1D4 antibody affinity column, which recognizes the 9 C‐terminal amino acids of rhodopsin tagged to the AE1 membrane domain. AE1‐Rho is non‐glycosylated and prodominantly dimeric, as determined by respectively endoglycosidase F treatment and size exclusion chromatography. AE1‐Rho secondary structure was assessed by circular dichroism. The functional activity of AE1‐Rho was measured by radioactive sulfate efflux assays in reconstituted proteoliposomes. This research is supported by the Canadian Institutes of Health Research.
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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 | 0.000 | 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".