The catalytic significance of the proposed active site residues in <i>Plasmodium falciparum</i> histoaspartic protease
Bibliographic record
Abstract
Alanine mutations of the proposed catalytically essential residues in histoaspartic protease (HAP) (H34A, S37A and D214A) were generated to investigate whether: (a) HAP is a serine protease with a catalytic triad of His34, Ser37 and Asp214 [Andreeva N, Bogdanovich P, Kashparov I, Popov M & Stengach M (2004) Proteins55, 705-710]; or (b) HAP is a novel protease with Asp214 acting as both the acid and the base during substrate catalysis with His34 providing critical stabilization [Bjelic S & Aqvist J (2004) Biochemistry43, 14521-14528]. Our results indicated that recombinant wild-type HAP, S37A and H34A were capable of autoactivation, whereas D214A was not. The inability of D214A to autoactivate highlighted the importance of Asp214 for catalysis. H34A and S37A mutants hydrolyzed synthetic substrate indicating that neither His34 nor Ser37 was essential for substrate catalysis. Both mutants did, however, have reduced catalytic efficiency (P < or = 0.05) compared with wild-type HAP, which was attributed to the stabilizing role of His34 and Ser37 during catalysis. The mature forms of wild-type HAP, H34A and S37A all exhibited high activity over a broad pH range of 5.0-8.5 with maximum activity occurring between pH 7.5 and 8.0. Inhibition studies indicated that wild-type HAP, H34A and S37A were strongly inhibited by the serine protease inhibitor phenylmethanesulfonyl fluoride, but only weakly inhibited by pepstatin A. The data, in concert with molecular modeling, suggest a novel mode of catalysis with a single aspartic acid residue performing both the acid and base roles.
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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".