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Benchmarking of Cph1 Mutants and DrBphP for Light-Responsive Phytochrome-Based Hydrogels with Reversibly Adjustable
Ramona Emig1,2,3,4, Philipp Hoess1, Hanyang Cai1
1Faculty of Biology, University of Freiburg, 79104, Freiburg, Germany.
Advanced Biology
|April 28, 2022
Summary
Engineered bacterial phytochromes enhance biohybrid hydrogels for optogenetics. A Cph1 mutant improves light-responsive stiffness, crucial for understanding cell mechanics and advancing optogenetic tools.
Area of Science:
- Molecular optogenetics
- Biomaterials science
- Cellular mechanobiology
Background:
- Optogenetic system performance depends on genetically encoded photoreceptor switching properties.
- Bacterial phytochromes are key light-sensitive proteins used in optogenetics.
Purpose of the Study:
- To engineer and characterize bacterial phytochromes (Cph1, DrBphP) for improved biohybrid hydrogel light responsiveness.
- To investigate the impact of modified hydrogel viscoelasticity on mammalian cell behavior.
Main Methods:
- Recombinant production of Cph1 and DrBphP in E. coli.
- Characterization of phytochrome switching properties.
- Synthesis and mechanical testing of biohybrid hydrogels.
- Stiffness measurements of human atrial fibroblasts on hydrogels.
Main Results:
- The Cph1 R472A mutant enhanced the dynamic range of hydrogel storage modulus.
- A distinct light-response in loss modulus was observed for the Cph1 R472A mutant hydrogel.
- Differences in hydrogel viscoelasticity influenced fibroblast stiffness, highlighting mechanotransduction importance.
Conclusions:
- Engineered phytochromes, particularly Cph1 R472A, advance biohybrid hydrogel light-responsive properties.
- Hydrogel viscoelasticity significantly impacts cellular mechanotransduction.
- These materials and findings are valuable for studying mammalian cell responses to dynamic matrix cues and for future optogenetic system development.
