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Facilities and methods for the high-throughput crystal structural analysis of human proteins
Udo Heinemann1, Konrad Büssow, Uwe Mueller
1Forschungsgruppe Kristallographie, Max-Delbrück-Centrum für Molekulare Medizin, Robert-Rössle-Strasse 10, D-13125 Berlin, Germany. Heinemann@MDC-Berlin.de
Accounts of Chemical Research
|March 19, 2003
Summary
The Protein Structure Factory streamlines human protein analysis using high-throughput methods. This structural genomics initiative employs recombinant expression, robotic crystallization, and synchrotron X-ray diffraction for rapid structure determination.
Area of Science:
- Structural biology
- Genomics
Background:
- High-throughput structural genomics projects aim to determine protein structures efficiently.
- Understanding human protein structures is crucial for biological and medical research.
Purpose of the Study:
- To describe the facilities and methods established at the Protein Structure Factory for high-throughput analysis of human protein crystal structures.
- To evaluate the strategy of the Protein Structure Factory based on initial results.
Main Methods:
- Recombinant expression of human proteins in Escherichia coli, Saccharomyces cerevisiae, or Pichia pastoris.
- Production of target proteins with affinity tags for standardized purification.
- Utilizing a robotic station for high-throughput crystallization (960,000 experiments capacity).
- X-ray diffraction experiments at the BESSY synchrotron storage ring.
Main Results:
- Establishment of facilities and methods for high-throughput human protein structure analysis.
- Implementation of recombinant expression, affinity purification, and robotic crystallization.
- Integration with synchrotron X-ray diffraction capabilities.
Conclusions:
- The Protein Structure Factory is operational, employing advanced technologies for structural genomics.
- The established strategy facilitates efficient production and structure determination of human proteins.
- Initial results support the viability and potential of the Protein Structure Factory's approach.