Results of JAXA-GCF Project --- High-quality Protein Crystallization under Microgravity for Structural Biology

Paper number

IAC-05-A2.3.01

Author

Mr. Sachiko Takahashi, Confocal Science, Japan

Coauthor

Dr. Koji Inaka, Japan

Coauthor

Mr. Masaru Sato, Japan Aerospace Exploration Agency (JAXA), Japan

Coauthor

Mr. Shinichi Shinozaki, Japan

Coauthor

Ms. Ari Yamanaka, Japan

Coauthor

Ms. Mari Yamanaka, Japan

Coauthor

Ms. Erika Hirota, Japan

Coauthor

Dr. Shigeru Sugiyama, Japan

Coauthor

Mr. Mitsuyasu Kato, Japan Aerospace Exploration Agency (JAXA), Japan

Coauthor

Ms. Chie Saito, Japan Aerospace Exploration Agency (JAXA), Japan

Coauthor

Mr. Satoshi Sano, Japan Aerospace Exploration Agency (JAXA), Japan

Coauthor

Mr. Moritoshi Motohara, Japan Aerospace Exploration Agency (JAXA), Japan

Coauthor

Mr. Tai Nakamura, Japan Aerospace Exploration Agency (JAXA), Japan

Coauthor

Mr. Tomoyuki Kobayashi, Japan Aerospace Exploration Agency (JAXA), Japan

Coauthor

Mr. Susumu Yoshitomi, Japan Aerospace Exploration Agency (JAXA), Japan

Year

2005

Abstract

Japan Aerospace Exploration Agency (JAXA) has been conducting the project for obtaining high-quality protein crystals under microgravity (JAXA-GCF project) twice a year since 2003, to contribute to the progress of structural biology using the space environment. In the project, we achieved a significant technical advancement, so that the success rate of the crystallization has become increased.
Alpha-amylase, a glycoprotein derived from Aspergillus oryzae, has been used as a technical verification protein for crystallization under microgravity. We obtained crystals of alpha-amylase which were diffracted beyond 0.89Å at SPring-8 beamline BL12B2 using polyethylene glycol (PEG) 8000 as a precipitant. Furthermore, crystals did not form cluster-like morphology which usually occurred on the ground experiment. The results indicated that microgravity effected on alpha-amylase crystals for improving crystal quality remarkably. From our numerical analysis, viscosity of the crystallization solution, caused by PEG, resulted in growing highly-ordered protein crystals for forming protein depletion zone around a crystal especially under microgravity. Based on this, we performed lysozyme crystallization experiment using NaCl as a precipitant in which PEG 8000 was added on purpose to increase viscosity of the crystallization solution to enhance the effects of microgravity. We successfully obtained lysozyme crystal which was diffracted 0.88Å at SPring-8 beamline BL12B2. The 0.89Å and 0.88Å resolution are among the highest resolution data in the Protein Data Bank to date.
We concluded that we have developed technologies for growing high-quality protein crystals in space for obtaining high resolution diffraction data, which will work out very effective for understanding 3-dimensional protein structures. 

Abstract document

IAC-05-A2.3.01.pdf

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