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J. Biochem, 2003, Vol. 133, No. 1 29-32
© 2003 Japanese Biochemical Society

BIOCHEMISTRY

Cloning, Expression, Crystallization, and Preliminary X-Ray Analysis of Recombinant Mouse Lipocalin-type Prostaglandin D Synthase, a Somnogen-Producing Enzyme

Daisuke Irikura+,1,2,3, Takashi Kumasaka§,3, Masaki Yamamoto3, Hideo Ago3, Masashi Miyano3, Kilunga Bruno Kubata2, Hiroaki Sakai4, Osamu Hayaishi2 and Yoshihiro Urade,1,2,3

1 Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, 6-2-4 Furuedai, Suita, Osaka 565-0874; 2 Department of Molecular Behavioral Biology, Osaka Bioscience Institute, 6-2-4 Furuedai, Suita, Osaka 565-0874; 3 RIKEN Harima Institute at SPring-8, 1-1-1 Kohto, Mikazuki-cho, Sayo, Hyogo 679-5148; and 4 Institute for Protein Research, Osaka University, Yamadaoka, Suita, Osaka 565-0871

Lipocalin-type prostaglandin D synthase is the key enzyme for the production of prostaglandin D2, a potent endogenous somnogen, in the brain. We cloned, produced, and crystallized the native enzyme and selenomethionyl Cys65Ala mutants of the recombinant mouse protein by the hanging drop vapor-diffusion method with both malonate and citrate as precipitants. The native crystals obtained with malonate belong to orthorhombic space group P212121 with lattice constants a = 46.2, b = 66.8, and c = 105.3 Å. The selenomethionyl crystals obtained with citrate belong to orthorhombic space group C2221 with lattice constants a = 45.5, b = 66.8, and c = 104.5 Å. The native crystals diffracted beyond 2.1 Å resolution.

+ Present address: Department of Medicine, Harvard Medical School, Division of Rheumatology, Immunology, and Allergy, Brigham and Women’s Hospital, Boston, MA 02115, USA.

§ Present address: Department of Life Science, Tokyo Insititute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501.

To whom correspondence should be addressed at: Department of Molecular Behavioral Biology, Osaka Bioscience Institute, 6-2-4 Furuedai, Suita, Osaka, 565-0874. Tel: +81-6-6872-4851, Fax: +81-6-6872-2841, E-mail: uradey{at}obi.or.jp


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