Oligomerization of Equilibrative Nucleoside Transporter 1
Bibliographic record
Abstract
Equilibrative nucleoside transporter 1 (ENT1) is a ubiquitously expressed membrane transporter in mammalian cells responsible for the transmembrane flux of endogenous nucleosides such as adenosine, as well as, chemotherapeutic, anti‐vial, and anti‐parasitic nucleoside analogues . The present study was designed to assess whether ENT1, like many other solute transporters, functions as a dimer or higher order oligomer. Previous functional studies from our laboratory suggested the presence of two populations of ENT1 with differential sensitivities to sulfhydryl reagents at the plasma membrane. Studies of ENT1 photoaffinity labelled membranes revealed multiple molecular mass bands on denaturing gels. Non‐denaturing Blue Native PAGE resulted in a molecular mass of ENT1 of 137 kDa (versus 55kDa monomer) suggesting a higher order oligomer or association with other proteins. Co‐immunoprecipitation studies on HEK293 cells expressing MYC‐ and HA‐hENT1 indicated that hENT1 can oligomerize. This was confirmed by immunofluorescence experiments, which showed a 1.5‐fold increase in colocalization of MYC‐hENT1 and HA‐hENT1 signal when the amount of transfected HA‐hENT1 cDNA was gradually increased while MYC‐hENT1 was kept constant in HEK293 cells. Finally, proximity ligation assays, performed on WT‐hENT1‐PK15 cells transiently transfected with MYC‐hENT1, showed interaction between the two pools of hENT1. These data support the hypothesis that ENT1 exists in the plasma membrane as a dimer, and possibly higher level protein complexes. Further work regarding the identity of the ENT1 complex components as well as the impact of these interactions on the function and inhibitor/substrate association with ENT1 remains to be determined. Supported by the Natural Science and Engineering Research Council of Canada.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| 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) | 0.002 | 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 source (direct Gemma or distilled Codex), 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".