Slabas A R, Cottingham I R, Austin A, Hellyer A, Safford R, Smith C G
Unilever Research, Sharnbrook, U.K.
Biochim Biophys Acta. 1990 Jun 19;1039(2):181-8. doi: 10.1016/0167-4838(90)90184-h.
An antibody has been raised against rape seed enoyl-ACP reductase. This recognizes both the alpha and beta polypeptides of the enzyme. Immunoblotting of fresh seed demonstrates that beta is not present in seed material, and that it is produced by proteolysis during isolation. It is thus deduced that rape seed enoyl reductase is an alpha 4 homotetramer. Leaf material from both rape and Arabidopsis have an enoyl reductase with a similar electrophoretic mobility to the rape seed enzyme when analyzed on SDS-PAGE. Quantitative immunoassay has demonstrated that the enzyme continually increases during lipid deposition, indicating that an increase in this enzyme is required to sustain high levels of lipid biosynthesis. In vitro translation experiments show that the enzyme is nuclear coded and synthesized as a precursor form. Immunogold electron microscopy has demonstrated that enoyl reductase is located in plastids. It is shown that ACP-Sepharose may be used as a matrix in the purification of enoyl-ACP reductase.
已经制备了一种针对油菜籽烯酰-ACP还原酶的抗体。该抗体可识别该酶的α和β多肽。对新鲜种子进行免疫印迹分析表明，种子材料中不存在β多肽，它是在分离过程中通过蛋白水解产生的。因此推断油菜籽烯酰还原酶是一种α4同四聚体。在SDS-PAGE上分析时，油菜和拟南芥的叶片材料中有一种烯酰还原酶，其电泳迁移率与油菜籽酶相似。定量免疫测定表明，该酶在脂质沉积过程中持续增加，这表明需要增加这种酶来维持高水平的脂质生物合成。体外翻译实验表明，该酶由细胞核编码并以前体形式合成。免疫金电子显微镜显示烯酰还原酶位于质体中。结果表明，ACP-琼脂糖可作为纯化烯酰-ACP还原酶的基质。
