Enzyme evolution by chemical complementationBaker, K.; Bleczinski, C.; Lin, H.; Salazar-Jimenez, G.; Sengupta, D.; Krane, S.; Cornish, V.W. Chemical complementation: a reaction-independent genetic assay for enzyme catalysis. Proc. Natl. Acad. Sci. USA 2002, 99 (26), 16537-16542.Azizi, B.; Chang, E.I.; Doyle, D.F. Chemical complementation: small-molecule-based genetic selection in yeast. Biochem. Biophys. Res. Commun.2003, 306, 774-780.Lin, H.; Tao, H.; Cornish, V.W. Directed evolution of a glycosynthasevia chemical complementation. J. Am. Chem. Soc. 2004, 126, 15051-15059.
Chemical complementation as developed in the Cornish lab•What are the authors trying to do?•How do they go about engineering their system of complementation?•What enzyme do they use to test their system? How does this work? (Figure 2, panel B)•What are the results? (Figure 3, panels B and C) How do the authors confirm that the change in transcription is caused by enzyme turnover?•Is this strategy general? What are the benefits and pitfalls?
This preview has intentionally blurred sections.
Sign up to view the full version.