title: DNA-directed control of enzyme-inhibitor complex formation: a modular approach to reversibly switch enzyme activity.
abstract: DNA-templated reversible assembly of an enzyme-inhibitor complex is presented as a new and highly modular approach to control enzyme activity. TEM1-β-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.
authors.affiliation: Laboratory of Chemical Biology and Institute for Complex Molecular Systems Eindhoven, University of Technology, Den Dolech 2, 5600 MB Eindhoven, The Netherlands.
authors.forename: Brian M G
authors.initials: BM
authors.lastname: Janssen
authors.affiliation: Laboratory of Chemical Biology and Institute for Complex Molecular Systems Eindhoven, University of Technology, Den Dolech 2, 5600 MB Eindhoven, The Netherlands.
authors.forename: Wouter
authors.initials: W
authors.lastname: Engelen
authors.affiliation: Laboratory of Chemical Biology and Institute for Complex Molecular Systems Eindhoven, University of Technology, Den Dolech 2, 5600 MB Eindhoven, The Netherlands.
authors.forename: Maarten
authors.initials: M
authors.lastname: Merkx
mesh_terms: D004247:DNA / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism*; D009841:Oligonucleotides / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D001618:beta-Lactamases / Q000235:genetics
chemical_list: D004791:Enzyme Inhibitors; D009097:Multienzyme Complexes; D009841:Oligonucleotides; D004247:DNA; D001618:beta-Lactamases
keywords: bionanotechnology; molecular switch; reporter enzyme; self-assembly; β-lactamase