Handicap-recover evolution leads to a chemically versatile, nucleophile-permissive protease
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Shafee, T., Gatti-Lafranconi, P., Minter, R., & Hollfelder, F. (2015). Handicap-recover evolution leads to a chemically versatile, nucleophile-permissive protease. ChemBioChem, 16 1866-1869. https://doi.org/10.1002/cbic.201500295
Mutation of the TEV protease nucleophile from cysteine to serine causes a ~10e4-fold activity loss. Ten rounds of directed evolution of the mutant, TEVSer, overcame the detrimental nucleophile exchange to recover near-wild-type activity in the mutant TEVSerX. Rather than respecialising TEV to the new nucleophile, all enzymes along the evolutionary trajectory retained the ability to also use the original cysteine nucleophile. Thus the adaptive evolution of TEVSer is paralleled by a neutral trajectory for TEVCys, in which mutations that increase serine nucleophile reactivity hardly affect the reactivity of cysteine. This apparent nucleophile permissiveness explains how nucleophile switches can occur in the phylogeny of the chymotrypsin-like protease PA superfamily. Despite changing the key component of its chemical mechanism, the evolved variants TEVSerX and TEVCysX have similar activities, which could potentially facilitate escape from adaptive conflict to enable active site evolution.
Protease, directed evolution, nucleophilic catalysis, adaptive protein evolution, catalytic promiscuity, PA clan, Tobacco Etch Virus protease
We acknowledge financial support from the Biotechnology and Biological Sciences Research Council and MedImmune. F. H. is an ERC Starting Investigator.
External DOI: https://doi.org/10.1002/cbic.201500295
This record's URL: https://www.repository.cam.ac.uk/handle/1810/248671
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Licence URL: http://creativecommons.org/licenses/by/4.0/