Inagaki T, Ohishi N, Tsukagoshi N, Ichihara C, Udaka S, Yagi K
Biochem Int. 1986 Dec;13(6):1045-50.
In vitro synthesis of D-amino acid oxidase [D-amino acid: O2 oxidoreductase (deaminating), EC 1.4.3.3], one of the peroxisomal flavin enzymes, was performed using a rabbit reticulocyte lysate system in order to elucidate the biosynthetic pathway of the enzyme. The apparent molecular weight of the synthesized enzyme protein was the same as that of D-amino acid oxidase purified from pig kidney. On the other hand, the enzyme protein was not detectable when a wheat germ lysate system was used for the translation. Denaturation of pig kidney poly(A)+ RNA with methylmercury hydroxide prior to the translation was found to enhance the synthesis of the enzyme protein. These results suggest a tight conformational structure of the mRNA used.
为阐明过氧化物酶体黄素酶之一的D-氨基酸氧化酶[D-氨基酸：O2氧化还原酶（脱氨基），EC 1.4.3.3]的生物合成途径，利用兔网织红细胞裂解液系统进行了该酶的体外合成。合成的酶蛋白的表观分子量与从猪肾中纯化的D-氨基酸氧化酶相同。另一方面，当使用小麦胚芽裂解液系统进行翻译时，未检测到该酶蛋白。发现在翻译前用氢氧化甲基汞使猪肾poly(A)+ RNA变性可增强该酶蛋白的合成。这些结果表明所用mRNA具有紧密的构象结构。
