Characterizing the structure and function of parkin co-regulated gene protein
Bibliographic record
Abstract
Parkin co-regulated gene protein (PACRG) is encoded 204 bp upstream of the Parkin gene, linked via a bi-directional promoter. Parkin is an ubiquitin ligase that regulates mitophagy and signaling pathways induced by various cellular stressors. Parkin loss-of-function mutations result in early-onset autosomal recessive Parkinson's disease (PD). First, PACRG is a component of Lewy bodies, a pathological hallmark of PD. Moreover, the quaking viable spontaneous mutant mouse, which displays neurological impairment, is a knockout of both Parkin and PACRG. Intriguingly, knockout of Parkin alone generates no apparent neurological phenotype, suggesting that PACRG has a neuroprotective role, perhaps functionally redundant with Parkin. Additionally, PACRG has been shown to generate and maintain the axoneme structure of flagella and cilia. Yet it is unclear how PACRG mediates these functions at the molecular level. Moreover, the three-dimensional structure of PACRG is unknown and it lacks known homologous domains. The objective of this thesis was to characterize the structure and molecular interactions of the PACRG protein. To this end, various expression and purification systems were tested. I have shown that full-length PACRG is unstable upon single expression in HEK293 and E. coli cells. However, deleting the non-conserved N-terminus residues generated large amounts of protein sequestered into inclusion bodies. After application of a refolding protocol, the C-terminal domain (conserved in all metazoans) bound directly to MEIG1, a protein necessary for spermiogenesis and axoneme formation. Co-expression with MEIG1 significantly increased PACRG protein yields and allowed the purification of full-length PACRG bound to MEIG1 in a 1:1 complex. This thesis thus provides a basis for future PACRG structural and functional studies. Notably, the production of recombinant PACRG will enable the study of its interactions with microtubules, thus providing insight into axoneme formation.
Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.
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) | 0.000 | 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 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".