Kaderbhai M A, Austen B M
Biochem J. 1984 Jan 1;217(1):145-57. doi: 10.1042/bj2170145.
The two-dimensional polyacrylamide-gel electrophoresis technique of O'Farrell [(1975) J. Biol. Chem 250, 4007-4021] was applied to resolve and analyse the polypeptide composition of dog pancreatic rough microsomal membranes, which were shown to be active in co-translational processing of preprolactin synthesized from pituitary mRNA in a translation system in vitro. About 100 polypeptides are resolved. Treatment of rough microsomal membranes with EDTA and high KCl concentration yielded membranes stripped of their ribosomes with retention of activity for translocation and processing. Stripped microsomal membranes showed a selective concentration of approximately 25 polypeptides in the membranes when analysed by two-dimensional polyacrylamide-gel electrophoresis. The two-dimensional electrophoretic profile was catalogued into polypeptides that are glycoproteins, those that contain free thiol groups disposed at the cytosolic surface of microsomal vesicles and those that are of secretory origin but have been entrapped in the microsomal preparation. Several secretory components, including amylase, procarboxypeptidases, lipase and anionic trypsinogen, were tentatively identified among the microsomal polypeptides. The rough and stripped microsomal membranes from dog pancreas show a characteristic set of seven major acidic polypeptides, which are also identifiable in microsomal-membrane preparations isolated from dog liver and rat liver. One of these polypeptides was identified as protein disulphide-isomerase (EC 5.3.4.1).
奥法雷尔的二维聚丙烯酰胺凝胶电泳技术[（1975年）《生物化学杂志》250，4007 - 4021]被用于解析和分析犬胰腺粗面微粒体膜的多肽组成，这些膜在体外翻译系统中对由垂体mRNA合成的前催乳素进行共翻译加工时表现出活性。大约分辨出了100种多肽。用EDTA和高浓度KCl处理粗面微粒体膜，得到了核糖体被去除但保留了转位和加工活性的膜。通过二维聚丙烯酰胺凝胶电泳分析，去除核糖体的微粒体膜在膜中显示出大约25种多肽的选择性富集。二维电泳图谱被分类为糖蛋白多肽、那些在微粒体小泡胞质表面含有游离巯基的多肽以及那些起源于分泌但被困在微粒体制备物中的多肽。在微粒体多肽中初步鉴定出了几种分泌成分，包括淀粉酶、羧肽酶原、脂肪酶和阴离子胰蛋白酶原。犬胰腺的粗面和去除核糖体的微粒体膜显示出一组特征性的七种主要酸性多肽，在从犬肝和大鼠肝分离的微粒体膜制备物中也可识别。其中一种多肽被鉴定为蛋白二硫键异构酶（EC 5.3.4.1）。
