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DNA Origami as Seeds for Promoting Protein Crystallization
Bo Zhang1, Andy R Mei2, Mark Antonin Isbell2
1Department of Chemistry , Renmin University of China , Beijing 100872 , P. R. China.
ACS Applied Materials & Interfaces
|November 29, 2018
Summary
DNA origami enhances protein crystallization probability. This novel seeding method, using folded M13 DNA scaffolds, significantly increases successful crystallization, especially for proteins like catalase at lower concentrations.
Area of Science:
- Biochemistry
- Nanotechnology
- Structural Biology
Background:
- Protein crystallization is crucial for structural determination but often challenging.
- Developing novel methods to improve crystallization success rates is essential for structural biology.
- DNA origami offers a programmable nanoscale platform with potential applications in biomolecular assembly.
Purpose of the Study:
- To investigate the efficacy of DNA origami as a seeding agent for protein crystallization.
- To determine if DNA origami can enhance the probability of successful protein crystallization.
- To explore the influence of DNA origami conformation on protein crystallization.
Main Methods:
- Construction of DNA origami using M13 DNA scaffolds and complementary staple strands.
- Addition of DNA origami (2-6 nM) to protein solutions (catalase, 1.0-7.0 mg/mL).
- Comparison of crystallization success rates between DNA origami-treated samples, non-folded DNA controls, and no-DNA controls.
Main Results:
- DNA origami significantly increased the proportion of successful protein crystallization, up to 11-fold.
- Crystallization enhancement was particularly notable for low protein concentrations (<5 mg/mL).
- Both examined DNA origami conformations (sheet and cylinder) improved crystallization compared to controls, with no significant difference between them.
Conclusions:
- DNA origami acts as an effective seed to promote protein crystallization.
- This method shows promise for improving crystallization yields, especially for challenging protein samples.
- The findings support the potential application of DNA origami in structural biology, particularly for proteins with limited availability or difficult crystallization properties.
