Nepokroeff C M, Porter J W
J Biol Chem. 1978 Apr 10;253(7):2279-83.
Fatty acid synthetase messenger RNA was obtained from rat liver polysomal RNA and then injected into Xenopus laevis oocytes. The radioactive fatty acid synthetase protein synthesized in the oocytes was identified by immunoprecipitation with anti-fatty acid synthetase antibody and the immunoprecipitate was then characterized by electrophoresis on sodium dodecyl sulfate-polyacrylamide gel. Co-migration of authentic fatty acid synthetase and the labeled product synthesized in oocytes was observed. Based on sucrose density gradient analysis, the rat liver fatty acid synthetase mRNA has a sedimentation coefficient of approximately 33 S, which agrees with the predicted minimum size necessary to code for the fatty acid synthetase protein. In addition, this mRNA was partially purified with oligo(dT)-cellulose, which indicates that it has a polyadenylate region. The relative in vivo rate of synthesis of fatty acid synthetase and the level of fatty acid synthetase mRNA in liver were also determined during the course of dietary induction of this enzyme. The results indicate that the dietary-induced increase in the level of fatty acid synthetase is probably due to an increased level of the fatty acid synthetase mRNA.
脂肪酸合成酶信使核糖核酸从大鼠肝脏多聚核糖体核糖核酸中获取,然后注入非洲爪蟾卵母细胞。卵母细胞中合成的放射性脂肪酸合成酶蛋白通过用抗脂肪酸合成酶抗体进行免疫沉淀来鉴定,接着通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳对免疫沉淀物进行表征。观察到天然脂肪酸合成酶与在卵母细胞中合成的标记产物共同迁移。基于蔗糖密度梯度分析,大鼠肝脏脂肪酸合成酶信使核糖核酸的沉降系数约为33S,这与编码脂肪酸合成酶蛋白所需的预测最小尺寸相符。此外,这种信使核糖核酸用寡聚(dT) - 纤维素进行了部分纯化,这表明它有一个聚腺苷酸区域。在饮食诱导该酶的过程中,还测定了肝脏中脂肪酸合成酶的相对体内合成速率以及脂肪酸合成酶信使核糖核酸的水平。结果表明,饮食诱导的脂肪酸合成酶水平升高可能是由于脂肪酸合成酶信使核糖核酸水平的增加。