Physico-chemical properties of recombinant AcCystatin, a cysteine protease inhibitor from Angiostrongylus cantonensis
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Bibliographic record
Abstract
Objective To express, purify, identify and analyze the physico-chemical properties of recombinant AcCystatin. Methods Recombinant AcCystatin was separated and purified through GST-resin affinity chromatography and digestion of thrombin. The soluble purified protein was identified by SDS-PAGE and amino acid sequencing , followed by physico-chemical properties study on ultraviolet spectroscopy, fluorescence spectroscopy, circular dichroism spectroscopy of AcCystatin and its inhibitory activity against human Cathepsin B , G , L and S. Results The purified AcCystatin with a molecular weight of 13 600, was detected through SDS-PAGE and the first 20 amino acid residues were identified by N-terminal protein sequencing (Edman degradation method). Anaylsis of AcCystatin ultraviolet spectrum, fluorescence spectrum and circular dichroism spectrum showed that AcCystatin contained a disulfide bond within the peptide chain , and the protein structure characterized by folding of the peptide chain into 3 types of secondary structures: alpha-helix (39.57%), beta-sheet (35.28%) and random coil (25.25%) . Furthermore , AcCystatin was demonstrated to possessed an obvious inhibitory activity against human Cathepsin B(IC50=57.04 nmol/L), L(IC50=89.28 nmol/L) and S(IC50=66.59 nmol/L), but no significant effect on Cathepsin G activity. Conclusion Based on previous research, we have obtained recombinant AcCystatin in high production, high purity, high solubility and strong biological effect, which provides basic parameter for further study on mechanism of immunoregulation of AcCystatin in angiostrongyliasis cantonensis.
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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) | 0.001 | 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 it