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Tail-to-side-chain cyclization stabilizes proteins on thermoresponsive ELP-based nanoparticles
Masayasu Mie1, Kai Fujiwara1, Makoto Ichikawa1
1Department of Life Science and Technology, School of Life Science and Technology, Institute of Science Tokyo, 4259 Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa 226-8501, Japan.
Genetically encoded cyclization enhances thermal stability of heat-labile proteins within thermoresponsive nanoparticle assemblies. This method protects proteins of interest (POIs) during self-assembly, improving activity retention at elevated temperatures.
Area of Science:
- Biotechnology
- Materials Science
- Protein Engineering
Background:
- Thermoresponsive elastin-like polypeptides (ELPs) form functional nanoparticles.
- High assembly temperatures can inactivate heat-labile proteins of interest (POIs).
Purpose of the Study:
- To develop a method for protecting POIs during ELP nanoparticle assembly.
- To enhance the thermal stability of POIs using genetically encoded cyclization.
Main Methods:
- Incorporated a SnoopTag/SnoopCatcher cyclization module into an ELP-poly(aspartic acid) (ELP-D) scaffold.
- Utilized Renilla luciferase (RLuc) as a model POI.
- Assessed thermal stability and nanoparticle characteristics before and after cyclization.
Main Results:
- Tail-to-side-chain cyclization increased melting and half-inactivation temperatures by ~6°C.
- ELP-D-cRLuc nanoparticles showed enhanced thermal stability without altering size or morphology.
- ELP-D-cRLuc nanoparticles retained ~70% activity after heating to 42°C, versus ~30% for non-cyclized constructs.
Conclusions:
- Genetically encoded cyclization effectively protects POIs during heat-triggered nanoparticle assembly.
- This approach enables improved activity retention of functional nanoparticles at elevated temperatures.
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