Commercial Protein Crystal Growth Experiment (CPCG-CVDA)

Objectives

The objective of this experiment were: (1) to produce large, well-ordered crystals of different proteins for use in x-ray diffraction studies and (2) to continue to develop the technology of microgravity protein crystal growth.

Shuttle-Mir Missions
STS-79, STS-91

Approach
Protein crystal growth was achieved through vapor diffusion using the CVDA hardware. This hardware consisted of 128 individual protein crystal experiments contained in a Commercial Refrigerator Incubator Module (CRIM). The experiments were conducted at 22 degrees Celsius. The macromolecular compounds chosen for flight experiments on STS-91 included Proteinase K, Lysozyme, Pseudomonal Protein A, NAD Synthetase, Clumping Protein-B, Multiple Adhesion Protein-1, Glyceraldehyde-3-Phosphate Dehydrogenase complexed to Inhibitor 1, Inhibitor 2, and Inhibitor 3, Grass Pollen Allergen Phlp 5b, MutT, and Neuraminidase. Following the Shuttle landing, the protein crystals were given to the respective investigators for post-mission x-ray diffraction analysis.

The Commercial Protein Crystal Growth Experiments on STS-79 were conducted in an experimental assembly consisting of 128 individual experiments housed in a Commercial Refrigerator/Incubator Module (CRIM) and stored in the Shuttle middeck. The proteins involved in these experiments were Trypsin Inhibitor, PEP Carboxykinase Complex, Van X, Van A, B, GDP Mannose Mannosyl Hydrolase, Grass Pollen Allergen Ph1 P5, Factor D, and Lysozyme (involved in experiments by two co-investigators). The experiments were loaded into the flight hardware at Cape Canaveral.

Results
Preliminary analysis indicates that the vapor diffusion experiments on STS-91 produced diffraction-quality crystals for eight of the twelve macromolecular compounds flown. Crystals of proteinase K, lysozyme, and the glyceraldehyde-3-phosphate dehydrogenase produced diffraction data that was superior to previously collected data.

Diffraction-quality crystals were obtained for five of the nine proteins flown on STS-79. X-ray diffraction data collected from these crystals was comparable to the best data obtained from the ground-grown crystals and was used to enhance and complete the x-ray data already collected for these compounds.

The protein crystal growth experiments were successful in producing diffraction-quality crystals for eight of the twelve compounds flown and, all aspects of the CVDA hardware performed well.

Earth Benefits
Knowledge gained in the protein crystal growth experiments contributed to the enhancement of the understanding of the three-dimensional structure of several proteins. This information will be used to advance our fundamental understanding of protein interactions and in structure-based drug design projects to find new drugs to treat diseases.

Publications
None available at this time.

Principal Investigators
Lawrence J. Delucas, O.D., Ph.D.
University of Alabama at Birmingham

Co-Investigators
Shigeo Aibara, Ph.D.
Christian Betzel, Ph.D.
Yancho Devedjiev, Ph.D.
Mark Jedrzedas, Ph.D.
Graeme Laver, Ph.D.
Glaucius Oliva, Ph.D.
Stephen Quirk, Ph.D.
Narayana Sthanam, Ph.D.
Wolfgang Weber, Ph.D.
Louis Delbaere, Ph.D.
James Knox, Ph.D.
Karen Moore, Ph.D.
Michael Spangfort, Ph.D.

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Page last updated: 07/16/1999

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