Characterization of C3adesArg/acylation stimulating protein (ASP) binding to C5L2 transfected cells and 3T3-L1 preadipocytes (94.17)
Bibliographic record
Abstract
Abstract C5L2 is a recently identified receptor for C5a and C3a/C3adesArg (ASP). C5a/C5adesArg binds C5L2 with high affinity with no functional activation. By contrast, some studies demonstrate C3a/ASP binding/activation of C5L2; others do not. We evaluated the influence of diverse ASP-C5L2 binding methods used. Orphan receptors (n>30) were screened with 125I-ASP (20oC & 4oC) or fluorescently labeled ASP (Fl-ASP, 37oC). Only C5L2 transfected cells were positive. Cell-associated Fl-ASP increased markedly from transiently-transfected < stably transfected < Fl-ASP sorted HEK-C5L2 for both hC5L2 and mC5L2. Similar results were obtained with transfected C5L2-CHO and C5L2-SW872 cells. Non-transfected cells ± Fl-ASP demonstrated background fluorescence only. Recombinant ASP prepared under non-denaturing conditions demonstrated 10X greater bioactivity vs. proteolytically-derived plasma ASP (maximal TG synthetic activity 100–300 nM vs. 5 uM). In adherent HEK-C5L2 (F-ASP sorted) and 3T3-L1 cells, blocking with 10% FCS, protamine sulfate or ovalbumin prevented 125I-ASP non-specific binding (NSB), while BSA blocking increased it. Optimal specific binding (Kd 66±14 nM) was obtained at 20oC (vs.4oC) in PBS or HAG-CM buffer. By contrast, cells in suspension demonstrated only non-specific binding. ASP-C5L2 binding has distinct characteristics that lead to phosphorylation, internalization and increased TG. Funding: CIHR
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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) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.004 | 0.002 |
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".