Computational structure-based redesign of enzyme activity. Export

@article{citeulike:4075036, abstract = {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.}, author = {Chen, C. Y. and Georgiev, I. and Anderson, A. C. and Donald, B. R.}, citeulike-article-id = {4075036}, citeulike-linkout-0 = {http://dx.doi.org/10.1073/pnas.0900266106}, citeulike-linkout-1 = {http://www.pnas.org/content/0900266106v1//.abstract}, citeulike-linkout-2 = {http://www.pnas.org/content/0900266106v1//.full.pdf}, citeulike-linkout-3 = {http://view.ncbi.nlm.nih.gov/pubmed/19228942}, citeulike-linkout-4 = {http://www.hubmed.org/display.cgi?uids=19228942}, day = {10}, doi = {10.1073/pnas.0900266106}, issn = {1091-6490}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, keywords = {design, enzyme, structure}, month = {March}, number = {10}, pages = {3764--3769}, posted-at = {2009-02-24 07:40:58}, priority = {2}, title = {Computational structure-based redesign of enzyme activity.}, url = {http://dx.doi.org/10.1073/pnas.0900266106}, volume = {106}, year = {2009} }

TY - JOUR ID - citeulike:4075036 L3 - citeulike-article-id:4075036 N2 - 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. IS - 10 JF - Proceedings of the National Academy of Sciences of the United States of America SN - 1091-6490 EP - 3769 TI - Computational structure-based redesign of enzyme activity. VL - 106 SP - 3764 KW - design KW - enzyme KW - structure AU - Chen, CY AU - Georgiev, I AU - Anderson, AC AU - Donald, BR PY - 2009/03/10/ UR - http://dx.doi.org/10.1073/pnas.0900266106 ER -