Altered expression of a quality control protease in E. coli reshapes the in vivo mutational landscape of a model enzyme.
| Autor: | Thompson S; Graduate Group in Biophysics, University of California San Francisco, San Francisco, United States., Zhang Y; Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, United States., Ingle C; The Green Center for Systems Biology, University of Texas Southwestern Medical Center, Dallas, United States., Reynolds KA; The Green Center for Systems Biology, University of Texas Southwestern Medical Center, Dallas, United States.; Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, United States., Kortemme T; Graduate Group in Biophysics, University of California San Francisco, San Francisco, United States.; Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, United States.; Chan Zuckerberg Biohub, San Francisco, United States. |
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| Jazyk: | angličtina |
| Zdroj: | ELife [Elife] 2020 Jul 23; Vol. 9. Date of Electronic Publication: 2020 Jul 23. |
| DOI: | 10.7554/eLife.53476 |
| Abstrakt: | Protein mutational landscapes are shaped by the cellular environment, but key factors and their quantitative effects are often unknown. Here we show that Lon, a quality control protease naturally absent in common E. coli expression strains, drastically reshapes the mutational landscape of the metabolic enzyme dihydrofolate reductase (DHFR). Selection under conditions that resolve highly active mutants reveals that 23.3% of all single point mutations in DHFR are advantageous in the absence of Lon, but advantageous mutations are largely suppressed when Lon is reintroduced. Protein stability measurements demonstrate extensive activity-stability tradeoffs for the advantageous mutants and provide a mechanistic explanation for Lon's widespread impact. Our findings suggest possibilities for tuning mutational landscapes by modulating the cellular environment, with implications for protein design and combatting antibiotic resistance. Competing Interests: ST, YZ, CI, KR, TK No competing interests declared (© 2020, Thompson et al.) |
| Databáze: | MEDLINE |
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