DNA-directed control of enzyme-inhibitor complex formation: a modular approach to reversibly switch enzyme activity

B.M.G. Janssen, W. Engelen, M. Merkx

Research output: Contribution to journalArticleAcademicpeer-review

21 Citations (Scopus)
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Abstract

DNA-templated reversible assembly of an enzyme–inhibitor complex is presented as a new and highly modular approach to control enzyme activity. TEM1-ß-lactamase and its inhibitor protein BLIP were conjugated to different oligonucleotides, resulting in enzyme inhibition in the presence of template strand. Formation of a rigid dsDNA linker upon addition of a complementary target strand disrupts the enzyme–inhibitor complex and results in the restoration of enzyme activity, enabling detection of as little as 2 fmol DNA. The noncovalent assembly of the complex allows easy tuning of target and template strands without changing the oligonucleotide-functionalized enzyme and inhibitor domains. Using a panel of eight different template sequences, restoration of enzyme activity was only observed in the presence of the target viral DNA sequence. The use of stable, well-characterized protein domains and the intrinsic modularity of our system should allow easy integration with DNA/RNA-based logic circuits for applications in biomedicine and molecular diagnostics.
Original languageEnglish
Pages (from-to)547-553
Number of pages7
JournalACS Synthetic Biology
Volume4
Issue number5
DOIs
Publication statusPublished - 2015

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