Szabó G, Marczinovits I, Komáromy L, Bajszar G, Molnár J
Mol Biol Rep. 1981 Aug 14;7(4):221-5. doi: 10.1007/BF00805756.
The isolation and characterization of HnRNP from cultured Drosophila melanogaster cells is described. HnRNP particles were extracted from the purified nuclei of sonication in the presence of rat liver cytosol RNAse inhibitor. The nuclear extract was centrifuged on a 15-30% sucrose gradient. The main part of the heterogeneous HnRNP material was localized in the 30 to 80S region of the sucrose gradient. According to the results of re-sedimentation studies the monomer particle was 45S. The buoyant density of HnRNP particles from different regions of the sucrose gradient were equal to approximately 1.4. The protein composition of the particles was analyzed by urea-SDS-polyacrylamide gel electrophoresis. There are five main and a few minor bands. Only the main polypeptides have a slightly higher molecular weight than those of the major polypeptides of 30S subparticles from rat liver nuclei. According to electron = microscopic studies the particles are heterogeneous and the average diameter was found to be 24-26 nm both on the basis of negative contrast and platinum-palladium shadowed pictures.
本文描述了从培养的果蝇细胞中分离和鉴定核不均一核糖核蛋白(HnRNP)的过程。在大鼠肝脏胞质核糖核酸酶抑制剂存在的情况下,从超声破碎后的纯化细胞核中提取HnRNP颗粒。将核提取物在15%-30%的蔗糖梯度上进行离心。不均一的HnRNP物质的主要部分位于蔗糖梯度的30至80S区域。根据再沉降研究结果,单体颗粒为45S。来自蔗糖梯度不同区域的HnRNP颗粒的浮力密度约为1.4。通过尿素-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析颗粒的蛋白质组成。有五条主要条带和几条次要条带。只有主要多肽的分子量略高于大鼠肝细胞核30S亚颗粒的主要多肽。根据电子显微镜研究,这些颗粒是不均一的,基于负染色和铂-钯阴影照片,发现平均直径为24-26nm。