Characterization of a common precursor to corticotropin and beta-lipotropin: cell-free synthesis of the precursor and identification of corticotropin peptides in the molecule.
Characterization of a common precursor to corticotropin and beta-lipotropin: cell-free synthesis of the precursor and identification of corticotropin peptides in the molecule.
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促肾上腺皮质激素和 β-促脂激素的常见前体的表征：前体的无细胞合成以及分子中促肾上腺皮质激素肽的鉴定。
DOI：
10.1073/pnas.74.11.4826
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发表时间：
1977
影响因子：
11.1
通讯作者：
Edward Herbert
中科院分区：
文献类型：
--
作者：
James L. Roberts;Edward Herbert
从AtT-20/D-16 v肿瘤细胞培养物中分离mRNA，并在mRNA依赖性网织红细胞无细胞系统中翻译。促肾上腺皮质激素（ACTH）产物通过双抗体免疫沉淀程序纯化，使用对ACTH的α（1-24）序列具有特异性的抗血清。通过十二烷基硫酸钠/凝胶电泳和胍-HCl柱上的凝胶过滤显示产物是均匀的，表观分子量（Mr）为28，500。当来自D-16 v细胞的膜结合多聚核糖体在网织红细胞无细胞系统中完成其新生链时，合成具有相同分子量的产物。从肿瘤细胞中分离的ACTH被分成三种不同表观Mr的蛋白质：29，000、32，000和34，000。无细胞产物含有与肿瘤细胞中合成的Mr 29，000 ACTH相同的赖氨酸、甲硫氨酸和苯丙氨酸标记的胰蛋白酶肽。胰蛋白酶肽分析还揭示了在Mr 28，500无细胞产物中存在α（1-39）序列，并表明在Mr 28，500分子中仅存在该序列的一个拷贝。
mRNA was isolated from cultures of AtT-20/D-16v tumor cells and translated in a mRNA-dependent reticulocyte cell-free system. The corticotropin (ACTH) product was purified by a double-antibody immunoprecipitation procedure using antisera specific for the alpha(1-24) sequence of ACTH. The product is shown by sodium dodecyl sulfate/gel electrophoresis and gel filtration on guanidine-HCl columns to be homogeneous with an apparent molecular weight (Mr) of 28,500. A product with the same molecular weight is synthesized when membrane-bound polysomes from D-16v cells are allowed to complete their nascent chains in a reticulocyte cell-free system. Mr 31,000 ACTH isolated from tumor cells has been separated into three proteins of different apparent Mr:29,000, 32,000, and 34,000. The cell-free product contains the same lysine-, methionine-, and phenylalanine-labeled tryptic peptides as the Mr 29,000 ACTH synthesized in the tumor cells. Tryptic peptide analysis also reveals the presence of the alpha(1-39) sequence in the Mr 28,500 cell-free product and suggests that there is only one copy of this sequence in the Mr 28,500 molecule.