DNA-directed control of enzyme-inhibitor complex formation: a modular approach to reversibly switch enzyme activity
| Autor: | Wouter Engelen, Bmg Brian Janssen, Maarten Merkx |
|---|---|
| Přispěvatelé: | Protein Engineering |
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
biology
Oligonucleotide Protein domain Oligonucleotides Biomedical Engineering DNA General Medicine Inhibitor protein Biochemistry Genetics and Molecular Biology (miscellaneous) beta-Lactamases Enzyme assay Protein Structure Tertiary chemistry.chemical_compound Biochemistry chemistry Multienzyme Complexes Enzyme inhibitor Coding strand biology.protein Enzyme Inhibitors Linker |
| Zdroj: | ACS Synthetic Biology, 4(5), 547-553. American Chemical Society |
| ISSN: | 2161-5063 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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