Optimizing Protein Stability In Vivo
| Autor: | Maximilian J. Kern, James Titchmarsh, Annekathrin von Hacht, Stuart L. Warriner, Lenz R. Steimer, James C.A. Bardwell, Sheena E. Radford, Linda Foit, Gareth J. Morgan |
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| Rok vydání: | 2009 |
| Předmět: |
Models
Molecular Protein Folding Mutant Biology Article Evolution Molecular 03 medical and health sciences In vivo Drug Resistance Bacterial Escherichia coli Selection Genetic Molecular Biology Gene 030304 developmental biology Genetics 0303 health sciences Protein Stability Escherichia coli Proteins 030302 biochemistry & molecular biology Cell Biology In vitro Colicin Biophysics Protein folding Chemical stability Carrier Proteins Function (biology) |
| Zdroj: | Molecular Cell. 36:861-871 |
| ISSN: | 1097-2765 |
| DOI: | 10.1016/j.molcel.2009.11.022 |
| Popis: | Identifying mutations that stabilize proteins is challenging because most substitutions are destabilizing. In addition to being of immense practical utility, the ability to evolve protein stability in vivo may indicate how evolution has formed today's protein sequences. Here we describe a genetic selection that directly links the in vivo stability of proteins to antibiotic resistance. It allows the identification of stabilizing mutations within proteins. The large majority of mutants selected for improved antibiotic resistance are stabilized both thermodynamically and kinetically, indicating that similar principles govern stability in vivo and in vitro. The approach requires no prior structural or functional knowledge and allows selection for stability without a need to maintain function. Mutations that enhance thermodynamic stability of the protein Im7 map overwhelmingly to surface residues involved in binding to colicin E7, implying that evolutionary pressures that drive Im7-E7 complex formation may have compromised the stability of the isolated Im7 protein. |
| Databáze: | OpenAIRE |
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