Reprogramming EF-hands for design of catalytically amplified lanthanide sensors

  • Korrie L. Mack
  • Olesia V. Moroz
  • Yurii S. Moroz
  • Alissa B. Olsen
  • Jaclyn M. McLaughlin
  • Ivan V. Korendovych
Original Paper


We recently reported that a computationally designed catalyst nicknamed AlleyCat facilitates C–H proton abstraction in Kemp elimination at neutral pH in a selective and calcium-dependent fashion by a factor of approximately 100,000 (Korendovych et al. in Proc. Natl. Acad. Sci. USA 108:6823, 2011). Kemp elimination produced a colored product that can be easily read out, thus making AlleyCat a catalytically amplified metal sensor for calcium. Here we report that metal-binding EF-hand motifs in AlleyCat could be redesigned to incorporate trivalent metal ions without significant loss of catalytic activity. Mutation of a single neutral residue at position 9 of each of the EF-hands to glutamate results in almost a two orders of magnitude improvement of selectivity for trivalent metal ions over calcium. Development of this new lanthanide-dependent switchable Kemp eliminase, named CuSeCat EE, provides the foundation for further selectivity improvement and broadening the scope of the repertoire of metals for sensing. A concerted effort in the design of switchable enzymes has many environmental, sensing, and metal ion tracking applications.


Protein design Metal sensor Lanthanides 



The authors thank Syracuse University for providing funds for this study, the Beckman Foundation for a scholarship for K.L.M., and Robert P. Doyle for use of his CD instrument.

Supplementary material

775_2013_985_MOESM1_ESM.pdf (5.1 mb)
Supplementary material 1 (PDF 5217 kb)


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Copyright information

© SBIC 2013

Authors and Affiliations

  • Korrie L. Mack
    • 1
  • Olesia V. Moroz
    • 1
  • Yurii S. Moroz
    • 1
  • Alissa B. Olsen
    • 1
  • Jaclyn M. McLaughlin
    • 1
  • Ivan V. Korendovych
    • 1
  1. 1.Department of ChemistrySyracuse UniversitySyracuseUSA

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