High-throughput identification of protein mutant stability computed from a double mutant fitness landscape.

ABSTRACT: The effect of a mutation on protein stability is traditionally measured by genetic construction, expression, purification, and physical analysis using low-throughput methods. This process is tedious and limits the number of mutants able to be examined in a single study. In contrast, functional fitness effects can be measured in a high-throughput manner by various deep mutational scanning tools. Using protein GB 1, we have recently demonstrated the feasibility of estimating the mutational stability effect ( ΔΔG) of single-substitution based on the functional fitness profile of all double-substitutions. The principle is to identify genetic backgrounds that have an exhausted stability margin. The functional effect of an additional substitution on these genetic backgrounds can then be used to

SUBMITTER: Wu NC 

PROVIDER: S-EPMC4815338 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature