Stabilizing proteins, simplified: A Rosetta-based webtool for predicting favorable mutations.
Rosetta molecular modeling program
molecular modeling
protein design
protein engineering
protein stability
Journal
Protein science : a publication of the Protein Society
ISSN: 1469-896X
Titre abrégé: Protein Sci
Pays: United States
ID NLM: 9211750
Informations de publication
Date de publication:
10 2022
10 2022
Historique:
revised:
06
08
2022
received:
18
06
2022
accepted:
21
08
2022
entrez:
29
9
2022
pubmed:
30
9
2022
medline:
1
10
2022
Statut:
ppublish
Résumé
Many proteins have low thermodynamic stability, which can lead to low expression yields and limit functionality in research, industrial and clinical settings. This article introduces two, web-based tools that use the high-resolution structure of a protein along with the Rosetta molecular modeling program to predict stabilizing mutations. The protocols were recently applied to three genetically and structurally distinct proteins and successfully predicted mutations that improved thermal stability and/or protein yield. In all three cases, combining the stabilizing mutations raised the protein unfolding temperatures by more than 20°C. The first protocol evaluates point mutations and can generate a site saturation mutagenesis heatmap. The second identifies mutation clusters around user-defined positions. Both applications only require a protein structure and are particularly valuable when a deep multiple sequence alignment is not available. These tools were created to simplify protein engineering and enable research that would otherwise be infeasible due to poor expression and stability of the native molecule.
Identifiants
pubmed: 36173174
doi: 10.1002/pro.4428
pmc: PMC9490798
doi:
Substances chimiques
Proteins
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
e4428Subventions
Organisme : NIGMS NIH HHS
ID : R35 GM131923
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35GM131923
Pays : United States
Informations de copyright
© 2022 The Protein Society.
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