High molecular weight forms of adrenocorticotropic hormone in the mouse pituitary and in a mouse pituitary tumor cell line.
High molecular weight forms of adrenocorticotropic hormone in the mouse pituitary and in a mouse pituitary tumor cell line.
复制标题
小鼠垂体和小鼠垂体肿瘤细胞系中高分子量形式的促肾上腺皮质激素。
作者：
B. Eipper;R. Mains
变性溶剂已被用来确定促肾上腺皮质激素（ACTH）活性的分子量在小鼠垂体中，在分泌ACTH的小鼠垂体瘤细胞系（AtT-20/D-16 v），并在组织培养基从垂体瘤细胞。ACTH活性通过放射免疫测定法和生物测定法进行定量。可以利用盐酸胍或十二烷基硫酸钠来表征多种形式的促肾上腺皮质激素，因为用这些变性剂处理猪促肾上腺皮质激素（促肾上腺皮质激素的39个氨基酸多肽形式，α（1-39））、垂体提取物、肿瘤细胞提取物和肿瘤细胞组织培养基不会降低免疫促肾上腺皮质激素活性。基于盐酸胍存在下的凝胶过滤，垂体瘤细胞和小鼠垂体的提取物含有三种不同分子量的ACTH活性。ACTH的主要形式具有类似于α（1-39）的分子量（分子量4000-5500），但存在显著量的两种更高分子量形式的ACTH：分子量6500-9000和分子量20，000 - 30，000。6500-9000分子量形式的ACTH是组织培养基中ACTH的主要形式;在培养基中没有α（1-39）大小的ACTH峰。在放射免疫测定中，所有三种形式的ACTH产生与猪α（1-39）平行的竞争性结合曲线;在生物测定（刺激小鼠肾上腺肿瘤细胞系中的类固醇生成）中，每种分子形式的ACTH的剂量反应曲线与猪α（1-39）平行。
Denaturing solvents have been used to determine the molecular weight of the adrenocorticotropic hormone (ACTH) activity in mouse pituitary, in an ACTH secreting mouse pituitary tumor cell line (AtT-20/D-16v), and in the tissue culture medium from the pituitary tumor cells. ACTH activity was quantitated by radioimmunoassay and by bioassay. It is possible to utilize guanidine hydrochloride or sodium dodecyl sulfate in characterizing the multiple forms of ACTH because treatment of porcine ACTH (the 39 amino acid polypeptide form of ACTH, alpha(1-39)), pituitary extracts, tumor cell extracts, and tumor cell tissue culture medium with these denaturants does not diminish the immunological ACTH activity. Based on gel filtration in the presence of guanidine hydrocholoride, extracts of the pituitary tumor cells and the mouse pituitary contain three distinct molecular weight classes of ACTH activity. The major form of ACTH has a molecular weight similar to alpha(1-39) (molecular weight 4000-5500), but there are significant amounts of two higher molecular weight forms of ACTH: molecular weight 6500-9000 and molecular weight 20,000-30,000. The 6500-9000 molecular weight form of ACTH is the major form of ACTH in the tissue culture medium; there is no peak of alpha(1-39) size ACTH in the medium. In the radioimmunoasay all three forms of ACTH generate competitive binding curves parallel to that of porcine alpha(1-39); in the bioassay (stimulation of steroidogenesis in a mouse adrenal tumor cell line) the dose response curve for each of the molecular forms of ACTH is parallel to that for porcine alpha(1-39).