Boileau G, Gossard F, Seidah N G, Chrétien M
Can J Biochem Cell Biol. 1983 Jun;61(6):333-9. doi: 10.1139/o83-046.
Polyadenylated RNA prepared from porcine pituitary intermediate lobes was translated in the cell-free rabbit reticulocyte system. Two translation products with apparent relative masses (Mrs) of 37 500 and 35 000 on polyacrylamide-sodium dodecyl sulfate gel electrophoresis were immunoprecipitated with a beta-melanophore-stimulating hormone (beta-MSH) antiserum. Tryptic digestions after citraconylation of the lysine residues and purification of the peptides by high performance liquid chromatography demonstrated that both polypeptides contained authentic beta-endorphin sequences. N-terminus microsequencing of the translation products labeled with [3H]leucine revealed the following partial amino acid sequence for the polypeptides with Mrs of 37 500 and 35 000: Leu 8, 11, 12, 13, 15, 16, 17, 20, 28, and 36 and Leu 3, 11, 12, 13, 15, 16, 17, 28, and 36, respectively. These results taken together prove the existence of two distinct mRNAs coding for proopiomelanocortin in the porcine pituitary intermediate lobe.
从猪垂体中间叶制备的聚腺苷酸化RNA在无细胞兔网织红细胞系统中进行翻译。在聚丙烯酰胺-十二烷基硫酸钠凝胶电泳上，两种表观相对分子质量（Mr）分别为37500和35000的翻译产物用β-促黑素细胞激素（β-MSH）抗血清进行免疫沉淀。赖氨酸残基经柠康酰化后进行胰蛋白酶消化，并通过高效液相色谱法纯化肽段，结果表明这两种多肽均含有真实的β-内啡肽序列。对用[3H]亮氨酸标记的翻译产物进行N端微测序，得到了相对分子质量为37500和35000的多肽的以下部分氨基酸序列：分别为Leu 8、11、12、13、15、16、17、20、28和36以及Leu 3、11、12、13、15、16、17、28和36。综合这些结果证明，猪垂体中间叶存在两种不同的编码阿片促黑激素皮质素原的mRNA。
