Computational structure-based redesign of enzyme activity.

Published

Journal Article

We report a computational, structure-based redesign of the phenylalanine adenylation domain of the nonribosomal peptide synthetase enzyme gramicidin S synthetase A (GrsA-PheA) for a set of noncognate substrates for which the wild-type enzyme has little or virtually no specificity. Experimental validation of a set of top-ranked computationally predicted enzyme mutants shows significant improvement in the specificity for the target substrates. We further present enhancements to the methodology for computational enzyme redesign that are experimentally shown to result in significant additional improvements in the target substrate specificity. The mutant with the highest activity for a noncognate substrate exhibits 1/6 of the wild-type enzyme/wild-type substrate activity, further confirming the feasibility of our computational approach. Our results suggest that structure-based protein design can identify active mutants different from those selected by evolution.

Full Text

Cited Authors

  • Chen, C-Y; Georgiev, I; Anderson, AC; Donald, BR

Published Date

  • March 2009

Published In

Volume / Issue

  • 106 / 10

Start / End Page

  • 3764 - 3769

PubMed ID

  • 19228942

Pubmed Central ID

  • 19228942

Electronic International Standard Serial Number (EISSN)

  • 1091-6490

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.0900266106

Language

  • eng