publication . Other literature type . Article . 2017

Trade-offs between enzyme fitness and solubility illuminated by deep mutational scanning.

Emily E. Wrenbeck; John-Paul Bacik; John-Paul Bacik; Ryszard Michalczyk; Timothy A. Whitehead; Justin R. Klesmith;
Open Access
  • Published: 14 Feb 2017
  • Publisher: Proceedings of the National Academy of Sciences
Abstract
<jats:p>Proteins are marginally stable, and an understanding of the sequence determinants for improved protein solubility is highly desired. For enzymes, it is well known that many mutations that increase protein solubility decrease catalytic activity. These competing effects frustrate efforts to design and engineer stable, active enzymes without laborious high-throughput activity screens. To address the trade-off between enzyme solubility and activity, we performed deep mutational scanning using two different screens/selections that purport to gauge protein solubility for two full-length enzymes. We assayed a TEM-1 beta-lactamase variant and levoglucosan kinase...
Subjects
free text keywords: Biological Sciences, Multidisciplinary, Solubility, High-throughput screening, Fitness landscape, Conserved sequence, Biochemistry, Mutation, medicine.disease_cause, medicine, Transport protein, Biology, Mutant, Active site, biology.protein
Funded by
NSF| CAREER: Programming proteins by deep sequencing and design
Project
  • Funder: National Science Foundation (NSF)
  • Project Code: 1254238
  • Funding stream: Directorate for Engineering | Division of Chemical, Bioengineering, Environmental, and Transport Systems
Abstract
<jats:p>Proteins are marginally stable, and an understanding of the sequence determinants for improved protein solubility is highly desired. For enzymes, it is well known that many mutations that increase protein solubility decrease catalytic activity. These competing effects frustrate efforts to design and engineer stable, active enzymes without laborious high-throughput activity screens. To address the trade-off between enzyme solubility and activity, we performed deep mutational scanning using two different screens/selections that purport to gauge protein solubility for two full-length enzymes. We assayed a TEM-1 beta-lactamase variant and levoglucosan kinase...
Subjects
free text keywords: Biological Sciences, Multidisciplinary, Solubility, High-throughput screening, Fitness landscape, Conserved sequence, Biochemistry, Mutation, medicine.disease_cause, medicine, Transport protein, Biology, Mutant, Active site, biology.protein
Funded by
NSF| CAREER: Programming proteins by deep sequencing and design
Project
  • Funder: National Science Foundation (NSF)
  • Project Code: 1254238
  • Funding stream: Directorate for Engineering | Division of Chemical, Bioengineering, Environmental, and Transport Systems
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