Functionally diverging molecular quasi-species evolve by crossing two enzymes
2006 (English)In: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 103, no 29, 10866-10870 p.Article in journal (Refereed) Published
Molecular evolution is frequently portrayed by structural relationships, but delineation of separate functional species is more elusive. We have generated enzyme variants by stochastic recombinations of DNA encoding two homologous detoxication enzymes, human glutathione transferases M1-1 and M2-2, and explored their catalytic versatilities. Sampled mutants were screened for activities with eight alternative substrates, and the activity fingerprints were subjected to principal component analysis. This phenotype characterization clearly identified at least three distributions of substrate selectivity, where one was orthogonal to those of the parent-like distributions. This approach to evolutionary data mining serves to identify emerging molecular quasi-species and indicates potential trajectories available for further protein evolution.
Place, publisher, year, edition, pages
2006. Vol. 103, no 29, 10866-10870 p.
directed evolution, DNA shuffling, glutathione transferase, library, multivariate analysis
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:uu:diva-97178DOI: 10.1073/pnas.0604030103ISI: 000239327200010PubMedID: 16829572OAI: oai:DiVA.org:uu-97178DiVA: diva2:172001