The Structure of a Thermophilic Kinase Shapes Fitness upon Random Circular Permutation.

ABSTRACT: Proteins can be engineered for synthetic biology through circular permutation, a sequence rearrangement in which native protein termini become linked and new termini are created elsewhere through backbone fission. However, it remains challenging to anticipate a protein's functional tolerance to circular permutation. Here, we describe new transposons for creating libraries of randomly circularly permuted proteins that minimize peptide additions at their termini, and we use transposase mutagenesis to study the tolerance of a thermophilic adenylate kinase (AK) to circular permutation. We find that libraries expressing permuted AKs with either short or long peptides amended to their N-terminus yield distinct sets of active variants and present evidence that this trend arises because permuted p

SUBMITTER: Jones AM 

PROVIDER: S-EPMC5122316 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature