<i>In Vivo</i> Formation of Protein Based Aqueous Microcompartments
Bibliographic record
Abstract
In this paper, we report the formation of protein based liquid droplets resulting in the formation of in vivo microcompartments in E. coli or tobacco cells. These microcompartments were generated by expressing elastin-like polypeptides (ELP), which have the ability to undergo a reversible phase transition, resulting in the formation of an aqueous two-phase system (ATPS) in the cytoplasm of the cell. We prove that these microcompartments are liquid by expressing a fusion protein consisting of ELP and GFP and by performing fluorescence recovery after photobleaching (FRAP) experiments at different stages of cell cultivation. In the initial phases of cell growth, the fusion protein concentration is low and is not sufficient to drive the formation of a second aqueous phase. As the intracellular fusion protein concentration increases with longer cultivation time, droplets start forming, and as protein expression continues, the droplets coalesce at the poles of the E. coli cells. FRAP experiments with cells at different growth stages reveals that the protein in these ELP based droplets is comprised of aqueous and not solid aggregates, as seen in typical inclusion bodies. Staining of the ribosomes and coimaging of the ELP-GFP fusion protein showed that these compartments exclude the protein making machinery of the cell, acting as depots for newly formed protein. It is also shown, in vitro, that ELP based droplets result in the exclusion of proteases, protecting proteins from degradation. Additional studies are still required to test this possibility in vivo. To the best of our knowledge, this is the first report characterizing the formation of an engineered extra aqueous phase in a living organism.
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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".