A Computational Library Design Protocol for Rapid Improvement of Protein Stability: FRESCO

OnderzoeksoutputAcademic

34 Citaten (Scopus)

Samenvatting

The ability to stabilize enzymes and other proteins has wide-ranging applications. Most protocols for enhancing enzyme stability require multiple rounds of high-throughput screening of mutant libraries and provide only modest improvements of stability. Here, we describe a computational library design protocol that can increase enzyme stability by 20–35 °C with little experimental screening, typically fewer than 200 variants. This protocol, termed FRESCO, scans the entire protein structure to identify stabilizing disulfide bonds and point mutations, explores their effect by molecular dynamics simulations, and provides mutant libraries with variants that have a good chance (>10%) to exhibit enhanced stability. After experimental verification, the most effective mutations are combined to produce highly robust enzymes.
Originele taal-2English
TitelProtein Engineering
SubtitelMethods and Protocols
RedacteurenN.J. Clifton
Plaats van productieNew York, NY
UitgeverijHumana Press
Pagina's69-85
Aantal pagina's7
Volume1685
ISBN van elektronische versie978-1-4939-7366-8
ISBN van geprinte versie978-1-4939-7364-4
DOI's
StatusPublished - 31-okt.-2017

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