Dores R M, Surprenant A
Gen Comp Endocrinol. 1984 Oct;56(1):90-9. doi: 10.1016/0016-6480(84)90065-0.
In order to investigate the biosynthesis of ACTH- and beta-endorphin-related substances in the pars distalis of Anolis carolinensis, explants of pars distali were incubated for 24 hr in a complete medium which contained [3H]tyrosine. Acid extracts of the incubates were immunoprecipitated with either an affinity-purified ACTH antiserum or an affinity-purified beta-endorphin antiserum and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Three distinct peaks of ACTH-related material were detected. The major peak comigrated with human ACTH(1-39), while two minor peaks corresponded in apparent molecular weight to ACTH biosynthetic intermediate and precursor-sized material. Three peaks of beta-endorphin-related material were also detected. The major peak comigrated with beta-endorphin(1-31), while two minor peaks corresponded to beta-lipotropin (LPH) and precursor-sized material. Sequential immunoprecipitation experiments indicated that the precursor-sized material had antigenic determinants for both ACTH and beta-endorphin. In addition this peak was identical in apparent molecular weight to the common precursor for alpha-melanocyte-stimulating hormone (alpha-MSH) and beta-endorphin in the pars intermedia of A. carolinensis (R.M. Dores, Peptides 3, 925-935). Analysis of extracts of reptile pars distalis by gel-filtration chromatography revealed a single peak of naloxone-reversible opiate bioactivity which coeluted with the peak of beta-endorphin-sized immunoreactivity. On a molar basis there is tenfold more opiate bioactivity in the reptile pars distalis than in the reptile pars intermedia.
为了研究卡罗来纳安乐蜥远侧部中促肾上腺皮质激素（ACTH）和β-内啡肽相关物质的生物合成，将远侧部的外植体在含有[3H]酪氨酸的完全培养基中孵育24小时。孵育物的酸提取物用亲和纯化的ACTH抗血清或亲和纯化的β-内啡肽抗血清进行免疫沉淀，并通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳进行分析。检测到三个不同的ACTH相关物质峰。主要峰与人类ACTH(1-39)迁移一致，而两个较小的峰在表观分子量上对应于ACTH生物合成中间体和前体大小的物质。还检测到三个β-内啡肽相关物质峰。主要峰与β-内啡肽(1-31)迁移一致，而两个较小的峰对应于β-促脂素（LPH）和前体大小的物质。连续免疫沉淀实验表明，前体大小的物质具有ACTH和β-内啡肽的抗原决定簇。此外，该峰在表观分子量上与卡罗来纳安乐蜥中间部中α-黑素细胞刺激素（α-MSH）和β-内啡肽的共同前体相同（R.M.多雷斯，《肽》3，925-935）。通过凝胶过滤色谱法分析爬行动物远侧部的提取物，发现一个纳洛酮可逆的阿片生物活性峰，该峰与β-内啡肽大小的免疫反应性峰共洗脱。以摩尔为基础，爬行动物远侧部中的阿片生物活性比爬行动物中间部中的高十倍。
