Controlled activation of protein rotational dynamics using smart hydrogel tethering

Area of Science:

• Biomaterials Science
• Protein Engineering
• Biophysics

Background:

• Stimulus-responsive hydrogels offer precise control over protein dynamics.
• Stabilizing proteins within hydrogels is crucial for various biotechnological applications.
• Understanding protein dynamics is key to harnessing their catalytic efficiencies.

Purpose of the Study:

• To develop a modular, stimulus-responsive hydrogel system for stabilizing and controlling protein dynamics.
• To investigate the impact of hydrogel formation and relaxation on protein structure and dynamics.
• To enable applications leveraging protein specificity and catalytic functions.

Main Methods:

• Engineered calmodulin (CaM) and poly(ethylene glycol) (PEG) matrix for modular hydrogel construction.
• Reversible protein tethering via an engineered CaM-binding sequence.
• Isotopic labeling ((13)C and (15)N) of maltose binding protein (MBP) for Nuclear Magnetic Resonance (NMR) spectroscopy.
• Utilized TROSY-HSQC NMR to measure protein dynamics and structural changes.

Main Results:

• Hydrogel formation suppressed protein dynamics and increased protein stability.
• Transient heating induced hydrogel matrix relaxation, enhancing protein dynamics.
• Observed resolution of substrate-induced large-amplitude domain rearrangements in proteins.
• Demonstrated successful stabilization and controlled release of functional proteins.

Conclusions:

• The developed hydrogel system effectively stabilizes proteins and controls their dynamics in a stimulus-responsive manner.
• This approach allows for the investigation of protein conformational changes and substrate interactions.
• The modular hydrogel platform holds promise for advanced applications in biocatalysis, biosensing, and drug delivery.