Allosteric Pathway Analysis — OHM + RINpy + DMS
Sits directly ON the catalytic relay. Glu side chain electrostatically tunes the catalytic acid Asp210. Highest allosteric signal among all beneficial mutations.
Negative as a single but validated in the ICCG multi-mutant. Proves allosteric compensation — the pathway mechanism rescues local fitness cost.
The ONLY position that is both allosteric handle AND DMS-beneficial. Pro rigidifies the loop, locking signal into a favorable conformation. 5/5 cross-tool consensus.
Mutations on different paths have orthogonal allosteric effects → low antagonistic epistasis risk.
Top candidate: Q217P + W104L + A207E + R286P + N140D