Nucleation-Supersaturation Dual-Drive Crystallization Strategy Enables Efficient Protein Crystallization

Area of Science:

• Biophysics
• Materials Science
• Crystallography

Background:

• Established protein crystallization methods face limitations due to single-point regulation of nucleation or supersaturation.
• Developing efficient strategies for high-quality protein crystal fabrication is crucial for structural biology and drug discovery.

Purpose of the Study:

• To develop a novel nucleation-supersaturation dual-driven crystallization (DDC) strategy for enhanced protein crystallization.
• To leverage dual surface and confinement effects for synergistic regulation of nucleation sites and solution supersaturation.

Main Methods:

• Utilized a p(PEGDA-co-DMAA) hydrogel template with pre-filled NaCl to create heterogeneous nucleation sites and control supersaturation.
• Applied the DDC strategy to hen egg white lysozyme (HEWL) and trypsin protein solutions.

Main Results:

• Achieved high-quality HEWL crystals with large sizes (100-300 µm) and well-defined morphologies (hexagon, tetragon).
• Demonstrated a 9-12 times faster nucleation time compared to the conventional hanging drop method.
• Successfully crystallized trypsin, showing versatility across different protein types and concentrations (10-50 mg mL⁻¹).

Conclusions:

• The DDC strategy offers significant advantages for efficient protein crystal fabrication.
• This method shows great potential for producing protein crystals suitable for diverse applications in structural biology and beyond.