Quantitation of rat liver messenger ribonucleic acid for malic enzyme during induction by thyroid hormone.
Quantitation of rat liver messenger ribonucleic acid for malic enzyme during induction by thyroid hormone.
复制标题
甲状腺激素诱导过程中大鼠肝脏信使核糖核酸苹果酸酶的定量。
DOI：
10.1021/bi00515a028
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发表时间：
1981
期刊：
影响因子：
2.9
通讯作者：
Oppenheimer,JH
中科院分区：
文献类型：
--
作者：
Towle,HC;Mariash,CN;Schwartz,HL;Oppenheimer,JH
霍华德C. Towle，* 卡里N.哈罗德·马里阿什Schwartz，and Jack H.奥本海默摘要：3，5，3 '-三碘-L-甲状腺原氨酸（T3）诱导大鼠肝胞浆苹果酸酶是由于编码这种蛋白质的信使核糖核酸（mRNA）水平的增加。为了探讨这种激素作用的机制，我们测量了T3和苹果酸酶mRNA之间的剂量-反应关系，以及T3给药和停药后苹果酸酶mRNA出现和消失的动力学。通过使用mRNA依赖的兔网织红细胞裂解物翻译系统定量编码苹果酸酶的信使RNA。翻译产物中苹果酸酶的鉴定是通过对NADP依赖酶的生物特异性吸附树脂--N（β-氨基己基）腺苷2 '，5'-二磷酸-琼脂糖进行层析，然后进行特异性免疫沉淀和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳来实现的。用这个程序，苹果酸酶mRNA的水平低至0.003%的总翻译产物在正常的，未诱导的大鼠肝脏可以测量。
Howard C. Towle,* Cary N. Mariash, Harold L. Schwartz, and Jack H. Oppenheimer abstract: The induction of cytosolic malic enzyme in rat liver by 3, 5, 3'-triiodo-L-thyronine (T3) is due to an increase in the level of messenger ribonucleic acid (mRNA) coding for this protein. To investigate the mechanism of this hormone action, we measured the dose-response relationship between T3 and malic enzyme mRNA and the kinetics of appearance and disappearance of malic enzyme mRNA following T3 admin-istration and withdrawal, respectively. Messenger RNA coding for malic enzyme was quantitated by using the mRNA-dependent rabbit reticulocyte lysate translational system. Identification of malic enzyme in the translational products was achieved by chromatography on A^-fb-aminohexyl) adenosine 2', 5'-diphosphate-agarose, a biospecific af-finity resin for NADP-dependent enzymes, followed by specific immunoprecipitation and sodium dodecyl sulfate-polyacryl-amide gel electrophoresis. With this procedure, levels of malic enzyme mRNA as low as 0.003% of the total translational products in normal, uninduced rat liver could be measured.