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Binding cofactors with triplex-based DNA motifs.

Christoph Kröner1, Anja Göckel, Wenjing Liu

  • 1Institut für Organische Chemie, Universität Stuttgart, 70569 Stuttgart (Germany), Fax: (+49) 711-685-64321.

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DNA binding motifs can store and release essential cofactors like NADH and SAM. Researchers demonstrated that altering DNA sequences tunes binding affinity and selectivity, with immobilized DNA beads releasing NADH at body temperature.

Keywords:
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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Synthetic Biology

Background:

  • Cofactors are crucial for cellular functions and biotechnological applications.
  • Developing methods to store and release cofactors is of significant interest.

Purpose of the Study:

  • To investigate the potential of triplex-based DNA binding motifs for cofactor storage and release.
  • To assess the binding affinity and selectivity of these DNA motifs for various cofactors.

Main Methods:

  • Design and synthesis of triplex-based DNA binding motifs.
  • Measurement of dissociation constants for cofactor binding (e.g., SAM, THF, NADH, FAD).
  • Sequence modification of DNA motifs to alter binding selectivity.
  • Immobilization of DNA motifs onto beads for cofactor storage and temperature-triggered release.

Main Results:

  • Triplex DNA motifs successfully bind a range of essential cofactors including NADH, FAD, SAM, acetyl CoA, and tetrahydrofolate (THF).
  • Measured dissociation constants ranged from 0.1 μM for SAM to 35 μM for THF.
  • DNA sequence modifications allowed for tuning of binding selectivity, differentiating between similar molecules like FAD and ATP.
  • Immobilized DNA beads demonstrated the capacity to store NADH and release it upon warming to body temperature.

Conclusions:

  • Triplex-based DNA motifs offer a versatile platform for the specific binding, storage, and controlled release of vital cofactors.
  • Sequence-dependent selectivity highlights the potential for designing custom DNA-based molecular storage systems.
  • DNA-functionalized beads represent a promising approach for developing cofactor delivery systems with temperature-responsive release capabilities.