Detection of Activated Rab7 GTPase with an Immobilized RILP Probe
Bibliographic record
Abstract
The dynamic and coordinated exchange of multiple GTPases between the cytosol and the phagosome membrane represents a critical process during phagosome biogenesis. In particular, acquisition of Rab7 is crucial for progression to the stage where formation of phagolysosomes is observed. Optimal Rab7 effector function requires its conversion to the GTP-bound form where it becomes activated. In light of this regulatory node, the GDP/GTP switch on the Rab7 molecule represents a tractable event to dissect the control of phagosome maturation by intracellular pathogen or their products. Direct measurement of Rab7 activation requires 32P-GTP binding to renatured Rab7 recovered by pull downs and resolved by SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and autoradiography. Here, we describe a novel, alternative, nonradioactive assay to measure Rab7 activity which takes advantage of the specific binding of activated (GTP bound) Rab7 to its effector RILP (Rab7 interacting lysosomal protein). Active Rab7 bound to immobilized recombinant RILP on latex beads can be detected quantitatively by either classical Western blotting or flow cytometry.
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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 | 0.001 | 0.001 |
| Meta-epidemiology (narrow) | 0.001 | 0.001 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.000 | 0.001 |
| Scholarly communication | 0.001 | 0.001 |
| Open science | 0.002 | 0.001 |
| Research integrity | 0.002 | 0.003 |
| Insufficient payload (model declined to judge) | 0.003 | 0.003 |
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".