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

Research output: Contribution to journalArticle

10 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)411-418
Number of pages8
JournalJournal of Biological Inorganic Chemistry
Volume18
Issue number4
DOIs
StatePublished - Apr 2013

Keywords

  • Lanthanides
  • Metal sensor
  • Protein design

ASJC Scopus subject areas

  • Biochemistry
  • Inorganic Chemistry

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