Cloning, expression, crystallization, and preliminary X-ray analysis of recombinant mouse lipocalin-type prostaglandin D synthase, a somnogen-producing enzyme
Cloning, expression, crystallization, and preliminary X-ray analysis of recombinant mouse lipocalin-type prostaglandin D synthase, a somnogen-producing enzyme
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DOI：
10.1093/jb/mvg006
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发表时间：
2003-01-01
影响因子：
2.7
通讯作者：
Urade, Y
中科院分区：
文献类型：
--
作者：
Irikura, D;Kumasaka, T;Urade, Y
Lipocalin型前列腺素D合成酶是在大脑中产生前列腺素D-2的关键酶，前列腺素D-2是一种强有力的内源性睡眠激素。我们以丙二酸和柠檬酸为沉淀剂，采用悬滴气相扩散法克隆、生产和结晶了重组小鼠蛋白的天然酶和硒蛋氨酰半胱氨酸突变体。用丙二酸酯合成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，c=105.3埃。用柠檬酸盐法合成的硒蛋氨酸晶体属于正交晶系C222(1)空间群，晶格常数a=45.5b=66.8，c=104.5埃。自然晶体的衍射率超过2.1埃分辨率。
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 Angstrom. 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 Angstrom. The native crystals diffracted beyond 2.1 Angstrom resolution.