A Single Mutation in a Regulatory Protein Produces Evolvable Allosterically Regulated Catalyst of Nonnatural Reaction†
Dr. Olesia V. Moroz
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorDr. Yurii S. Moroz
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Department of Chemistry, Taras Shevchenko National University of Kiev, 64 Volodymyrska St., Kiev, 01601 (Ukraine)
Search for more papers by this authorDr. Yibing Wu
Department of Pharmaceutical Chemistry, University of California, San Francisco, 555 Mission Bay Blvd. South, San Francisco, CA 94158 (USA)
Search for more papers by this authorAlissa B. Olsen
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorDr. Hong Cheng
Fox Chase Cancer Center, 333 Cottman Ave., Philadelphia, PA 19111 (USA)
Search for more papers by this authorKorrie L. Mack
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorJaclyn M. McLaughlin
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorElizabeth A. Raymond
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorKrystyna Zhezherya
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorProf. Dr. Heinrich Roder
Fox Chase Cancer Center, 333 Cottman Ave., Philadelphia, PA 19111 (USA)
Search for more papers by this authorCorresponding Author
Prof. Dr. Ivan V. Korendovych
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)Search for more papers by this authorDr. Olesia V. Moroz
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorDr. Yurii S. Moroz
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Department of Chemistry, Taras Shevchenko National University of Kiev, 64 Volodymyrska St., Kiev, 01601 (Ukraine)
Search for more papers by this authorDr. Yibing Wu
Department of Pharmaceutical Chemistry, University of California, San Francisco, 555 Mission Bay Blvd. South, San Francisco, CA 94158 (USA)
Search for more papers by this authorAlissa B. Olsen
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorDr. Hong Cheng
Fox Chase Cancer Center, 333 Cottman Ave., Philadelphia, PA 19111 (USA)
Search for more papers by this authorKorrie L. Mack
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorJaclyn M. McLaughlin
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorElizabeth A. Raymond
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorKrystyna Zhezherya
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Search for more papers by this authorProf. Dr. Heinrich Roder
Fox Chase Cancer Center, 333 Cottman Ave., Philadelphia, PA 19111 (USA)
Search for more papers by this authorCorresponding Author
Prof. Dr. Ivan V. Korendovych
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, NY 13244 (USA)Search for more papers by this authorWe thank Prof. Robert P. Doyle for providing access to CD instrument. The Spectroscopy Support Facility at FCCC provided access to a 600 MHz NMR spectrometer.
Graphical Abstract
It only takes one mutation: A strategically placed single mutation in a non-enzymatic protein scaffold produced AlleyCat, a small, allosterically regulated catalyst of Kemp elimination. In only seven rounds of directed evolution the enzymatic efficiency of the original 74 amino acid residue catalyst was improved more than 220-fold to achieve a kcat value higher than that of catalytic antibodies for the same reaction, still preserving allosteric regulation.
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