Irikura Daisuke, Kumasaka Takashi, Yamamoto Masaki, Ago Hideo, Miyano Masashi, Kubata Kilunga Bruno, Sakai Hiroaki, Hayaishi Osamu, Urade Yoshihiro
Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, 6-2-4 Furuedai, Suita, Osaka 565-0874.
J Biochem. 2003 Jan;133(1):29-32. doi: 10.1093/jb/mvg006.
Lipocalin-type prostaglandin D synthase is the key enzyme for the production of prostaglandin D(2), a potent endogenous somnogen, in the brain. We cloned, produced, and crystallized the native enzyme and selenomethionyl Cys(65)Ala 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 P2(1)2(1)2(1) with lattice constants a = 46.2, b = 66.8, and c = 105.3 A. The selenomethionyl crystals obtained with citrate belong to orthorhombic space group C222(1) with lattice constants a = 45.5, b = 66.8, and c = 104.5 A. The native crystals diffracted beyond 2.1 A resolution.
脂联素型前列腺素D合成酶是大脑中产生前列腺素D2（一种强效内源性促眠物质）的关键酶。我们通过悬滴气相扩散法，以丙二酸盐和柠檬酸盐作为沉淀剂，克隆、制备并结晶了重组小鼠蛋白的天然酶和硒代蛋氨酸取代半胱氨酸65的丙氨酸突变体。用丙二酸盐获得的天然晶体属于正交晶系空间群P2(1)2(1)2(1)，晶格常数a = 46.2、b = 66.8和c = 105.3 Å。用柠檬酸盐获得的硒代蛋氨酸晶体属于正交晶系空间群C222(1)，晶格常数a = 45.5、b = 66.8和c = 104.5 Å。天然晶体的衍射分辨率超过2.1 Å。
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