Vindrola O, Ase A, Aloyz R, Saravia F, Finkielman S, Nahmod V E
Instituto de Investigaciones Médicas, Sección Sustancias Vasoactivas, Buenos Aires, Argentina.
J Mol Endocrinol. 1990 Oct;5(2):175-83. doi: 10.1677/jme.0.0050175.
Proenkephalin-derived peptides, in common with other prohormones, are associated with membranes of microsomes and secretory granules in the bovine adrenal medulla. Post-translational processing of the precursor molecule varies depending upon the tissue. The relationship between post-translational events in different tissues was examined by studying the membrane association of endogenous proenkephalin-derived peptides in the crude microsomal fraction of rat adrenal medulla, brain striatum and heart ventricle. [Met]-Enkephalin and synenkephalin (proenkephalin(1-70)) immunoreactivities were quantified by radioimmunoassay after sequential enzymatic digestion with trypsin and carboxypeptidase B. Between 60 and 75% of total immunoreactive peptides present in intact microsomes of the three tissues were associated with membranes and specifically released with 2 M KSCN (pH 7.4). Analysis of the chromatographic profile of materials present in the soluble and associated fractions produced the following results. In the three tissues the materials associated with microsomal membranes corresponded to peptides larger than 3-5 kDa and displayed synenkephalin and [Met]-enkephalin immunoreactivity. Adrenal and heart microsomes showed a continuous pattern of membrane-associated proenkephalin-derived peptides of high, intermediate and low molecular weights containing the synenkephalin and [Met]-enkephalin sequences. These tissues, however, presented quantitative differences, as the highest concentrations belonged to materials larger and smaller than 12.5 kDa in adrenal and heart microsomes respectively. On the other hand, brain striatal microsomes displayed a discontinuous pattern of associated materials, with the absence of some products of high and intermediate molecular weight. Only in the soluble fraction of striatal microsomes were peptides detected of high and intermediate molecular weight containing the [Met]-enkephalin but not the synenkephalin sequence.(ABSTRACT TRUNCATED AT 250 WORDS)
脑啡肽原衍生肽与其他激素原一样,与牛肾上腺髓质中的微粒体膜和分泌颗粒膜相关。前体分子的翻译后加工因组织而异。通过研究大鼠肾上腺髓质、脑纹状体和心室粗微粒体部分中内源性脑啡肽原衍生肽的膜结合情况,来检测不同组织中翻译后事件之间的关系。在用胰蛋白酶和羧肽酶B进行顺序酶消化后,通过放射免疫测定法定量[甲硫氨酸] - 脑啡肽和脑啡肽原(脑啡肽原(1 - 70))的免疫反应性。在这三种组织完整微粒体中存在的总免疫反应性肽中,60%至75%与膜相关,并在用2M KSCN(pH 7.4)处理时特异性释放。对可溶性和相关部分中存在的物质进行色谱分析得到以下结果。在这三种组织中,与微粒体膜相关的物质对应于大于3 - 5 kDa的肽,并显示出脑啡肽原和[甲硫氨酸] - 脑啡肽免疫反应性。肾上腺和心脏微粒体显示出高分子量、中分子量和低分子量的与膜相关的脑啡肽原衍生肽的连续模式,包含脑啡肽原和[甲硫氨酸] - 脑啡肽序列。然而,这些组织存在定量差异,因为肾上腺和心脏微粒体中最高浓度分别属于大于和小于12.5 kDa的物质。另一方面,脑纹状体微粒体显示出相关物质的不连续模式,缺少一些高分子量和中分子量的产物。仅在纹状体微粒体的可溶性部分中检测到含有[甲硫氨酸] - 脑啡肽但不含有脑啡肽原序列的高分子量和中分子量肽。(摘要截于250字)