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Crystallizing proteins - a rational approach?
1Department of Pharmaceutical Research-New Technologies, F. Hoffmann-La Roche Ltd., Basel, Switzerland.
Summary
Recombinant DNA technology enables large-scale pure protein production for structural studies. Advances in protein crystallization techniques and robotic screening accelerate the determination of three-dimensional protein structures.
Area of Science:
- Biochemistry and structural biology
- Molecular biology and genetic engineering
Background:
- Recombinant DNA technology has revolutionized protein production, yielding large quantities of pure proteins.
- This advancement overcomes limitations of natural protein sources for crystallization experiments.
- Increased involvement of laboratories in protein crystallization generates novel techniques to overcome challenges.
Purpose of the Study:
- To highlight the impact of recombinant DNA technology on protein crystallization.
- To discuss methods for overcoming the 'bottleneck of crystallization' in structural biology.
- To outline criteria and strategies for successful protein crystallization.
Main Methods:
- Utilizing large amounts of pure protein produced via recombinant DNA technology.
- Applying new techniques to address crystallization challenges.
- Employing robotic screening for optimizing crystallization conditions.
- Evaluating protein properties like solubility, purity, and aggregation.
Main Results:
- Facilitation of previously impossible experiments due to ample pure protein availability.
- Generation of new information and techniques to improve crystallization success rates.
- Development of strategies for protein modification to enhance crystal formation.
Conclusions:
- Recombinant DNA technology is pivotal for advancing protein structure determination.
- Robotic screening and pre-crystallization assessments improve efficiency.
- Protein engineering offers solutions for recalcitrant crystallization cases.