JBIC Journal of Biological Inorganic Chemistry

, Volume 18, Issue 4, pp 411–418

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

DOI: 10.1007/s00775-013-0985-5

Cite this article as:
Mack, K.L., Moroz, O.V., Moroz, Y.S. et al. J Biol Inorg Chem (2013) 18: 411. doi:10.1007/s00775-013-0985-5

Abstract

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.

Keywords

Protein design Metal sensor Lanthanides 

Supplementary material

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

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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