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Combining Wet and Dry Lab Techniques to Guide the Crystallization of Large Coiled-coil Containing Proteins
Published on: January 6, 2017
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Tuning coiled coil stability with histidine-metal coordination
Isabell Tunn1, Alberto S de Léon, Kerstin G Blank
1Max Planck Institute of Colloids and Interfaces, Science Park Potsdam-Golm, 14424 Potsdam, Germany. Kerstin.Blank@mpikg.mpg.de.
Nanoscale
|December 1, 2018
Summary
Researchers engineered synthetic coiled coils (CCs) with metal coordination sites. This enhances CC stability and mechanical reinforcement, improving biomimetic hydrogel properties.
Area of Science:
- Biomaterials Science
- Nanotechnology
- Biochemistry
Background:
- Coiled coils (CCs) are essential protein structures used in nanostructures, drug delivery, and hydrogels.
- Engineering CCs with tunable properties is crucial for advanced biomaterials.
Purpose of the Study:
- To create a nanoscale building block with tunable stability by incorporating metal coordination sites into synthetic CCs.
- To investigate the effect of metal ion binding on CC stability and mechanical properties.
- To assess the impact of these engineered CCs on the bulk properties of hydrogels.
Main Methods:
- Bioengineering synthetic coiled coils with terminal metal coordination sites.
- Circular dichroism (CD) spectroscopy to assess thermodynamic stability.
- Atomic force microscopy (AFM)-based single-molecule force spectroscopy to measure mechanical reinforcement.
- Incorporation of engineered CCs as dynamic crosslinks in poly(ethylene glycol) (PEG) hydrogels.
Main Results:
- Reversible Ni2+ coordination was shown to thermodynamically stabilize the CC.
- Ni2+-binding significantly reinforced the CC, increasing its dissociation energy barrier.
- Single-molecule stability of the CC translated to bulk hydrogel properties, influencing viscoelasticity.
Conclusions:
- Bioengineering metal coordination sites into CCs provides a strategy for tunable stability and mechanical reinforcement.
- This approach effectively engineers single-molecule properties of biomolecular building blocks.
- The tunable CCs can be translated to control emergent properties of biomimetic materials and molecular assemblies.
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