Three further constraint tests. Each asks the same question in a different field: does the claimed effect survive when the parameters are tied to something measurable rather than left free?
If tracking error depends only on R = τ/T, one curve should describe every system. At R = 1 the error spans 0.702 to 1.919 — a factor 2.74. Against the first-order curve the worst departure is 1.26 in absolute error amplitude.
| System | error at R = 1 |
|---|---|
| 1st order | 0.7016 |
| 2nd zeta=0.3 | 1.9193 |
| 2nd zeta=0.7 | 1.2223 |
| 2nd zeta=1.0 | 1.1145 |
| 2nd zeta=2.0 | 1.0298 |
Verdict: not supported. R is the dimensionless frequency; it was never going to carry order and damping. The hypothesis was a restatement, and the restatement does not extend.
The competitive-kinetics crossover is already fixed by kcat/Km; the only testable content is a departure. A synthetic two-site system was generated (clearly labelled synthetic), then fitted with a one-site model in two ways:
| Fit | RMSE | Meaning |
|---|---|---|
| one-site, Km free | 0.020 | the model absorbs the second site into a shifted Km |
| one-site, Km constrained to measured value | 0.180 | the departure becomes visible |
The constrained model is 9.0× worse than the free one. That is the test working as designed: when the free fit is much better than the constrained fit, the system is not described by single-site competitive kinetics, and the size of the gap measures how far off it is.
But this is a synthetic demonstration, not evidence about any real enzyme. Its only content is that the test has power when the departure exists. Whether real enzymes show it is untested and requires the measured constants.
The claim was that the optimum spacing-to-droplet ratio moves when collection and drainage are both scored. In the illustrative model used here, interception rises with spacing while the drainage fraction falls, and the product is maximised at the same spacing as interception alone — a shift factor of 1.00, i.e. no shift.
This test is uninformative as constructed, and the failure is mine, not the hypothesis's. The drainage term was monotone in the wrong direction, so no interior optimum could form. A useful version needs a drainage model with a genuine maximum (shedding that improves with spacing up to a point, then degrades), which was not built. The hypothesis remains planned, not refuted.
Applied three times on this page, the protocol produced one clean refutation, one synthetic demonstration of test power, and one honest construction failure. That mix is what a working protocol looks like.
| # | Hypothesis | Test |
|---|---|---|
| P3-H1 | Two-parameter collapse (R, ζ) is minimal for second-order systems; no one-parameter version exists for order ≥ 2. | Fit surfaces; held-out prediction across unseen ζ and order. |
| P3-H2 | Measured enzyme pairs will show constrained-fit residuals systematically larger than free-fit residuals, and the gap will track known allostery. | Use BRENDA/SABIO-RK constants; compare to allostery annotations. |
| P3-H3 | A drainage model with a non-monotone shedding term produces an interior fog optimum that monotone drainage cannot. | Build the non-monotone term; map the optimum. |
Status: computational model results, 3 November 2026. No experiment, no data, no measurement. No novelty claimed; no literature search performed. Code and raw JSON accompany this page set.