Nuclear envelope glycoprotein with poly(A) polymerase activity of rat liver: isolation, characterization, and immunohistochemical localization.
Nuclear envelope glycoprotein with poly(A) polymerase activity of rat liver: isolation, characterization, and immunohistochemical localization.
复制标题
具有大鼠肝脏聚腺苷酸聚合酶活性的核膜糖蛋白：分离、表征和免疫组织化学定位。
DOI：
10.1021/bi00425a015
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发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
Sidransky,H
中科院分区：
文献类型：
--
作者：
Kurl,RN;Holmes,SC;Verney,E;Sidransky,H
乔治华盛顿大学医学中心病理学系，华盛顿特区 20037 收稿日期：1988 年 3 月 25 日；修订稿于 1988 年 7 月 28 日收到摘要：已鉴定出一种具有多聚 (A) 聚合酶活性的蛋白质，并从大鼠的肝核包膜中分离出接近同质的蛋白质。该酶与伴刀豆球蛋白 A-琼脂糖结合并用甲基 α-D-吡喃甘露糖苷 (0.2 M) 从柱上洗脱的能力以及 α-甘露糖苷酶的抑制作用表明它是一种糖蛋白。 Poly (A) 聚合酶绝对需要二价阳离子、ATP 和寡核苷酸引物。 Mn2+酶活比Mg2+酶活高约20倍。几种已知的抑制剂会对聚腺苷酸聚合酶活性产生不利影响。在变性条件下通过聚丙烯酰胺凝胶电泳分析时，该酶的分子量为 64 000，沉降系数为 4.5 S。使用针对纯化酶产生的多克隆抗体进行的免疫组织化学研究表明，该抗原位于核膜中。^^^ 腺苷酸化是 hnRNA 在作为 mRNA 转运到细胞质之前参与成熟的核过程之一。 Poly (A) 序列已涉及这项工作得到了国家癌症研究所的美国公共卫生服务研究补助金 CA-41832 和国家关节炎、糖尿病、消化和肾脏疾病研究所的 DK-27339 的支持。 (i) mRNA 稳定性（Nudel 等人，1976），(ii) mRNA 的核细胞质转运（Muller 等人，1984），以及（iii）蛋白质合成（Jacobson & Favreau，1983；Rubin & Halim，1987）。然而，大多数组蛋白 mRNA 缺乏多聚 (A) 序列（Adesnik & Darnell，1972；Greenberg & Perry，1972），而且最近从成年大鼠大脑中分离出的 mRNA 群体也被证明缺乏该序列（Chaudhari & Hahn，1983）。
Department of Pathology, The George Washington University Medical Center, Washington, DC 20037 Received March 25, 1988; Revised Manuscript Received July 28, 1988 abstract: A protein with poly (A) polymerase activity has been identified and isolated from hepatic nuclear envelopes of rats to near homogeneity. The ability of the enzyme to bind to concanavalin A-agarose and to be eluted from the column with methyl aD-mannopyranoside (0.2 M) as well as the inhibitory effects of a-mannosidase suggested that it was a glycoprotein. Poly (A) polymerase has an absolute requirement for a divalent cation, ATP, and an oligonucleotide primer. The enzyme activitywith Mn2+ was about 20-fold higher than thatwith Mg2+. Several known inhibitors adversely affected poly (A) polymerase activity. The enzyme has a molecular weight of 64 000 when analyzedby polyacrylamide gel electrophoresis under denaturing conditions and has a sedimentation coefficient of 4.5 S. Immunohistochemical studies using polyclonal antibodies raised against the purified enzyme revealed that the antigen was localized in the nuclear membranes.^^^ ilyadenylation is one of the nuclear processes involved in the maturation of hnRNA prior to its transport to the cyto-plasm as mRNA. Poly (A) sequences have been implicated tThis work was supported by US Public Health Service Research Grants CA-41832 from the National Cancer Institute and DK-27339 from the National Institute of Arthritis, Diabetes, and Digestive and Kidney Diseases. in (i) mRNA stability (Nudel et al., 1976),(ii) nucleocytoplasmic transport of mRNA (Muller et al., 1984), and (iii) protein synthesis (Jacobson & Favreau, 1983; Rubin & Halim, 1987). However, most histone mRNAs lack poly (A) se-quences (Adesnik & Darnell, 1972; Greenberg & Perry, 1972), and also a population of mRNA isolated from adult rat brains has recently been shown to lack the sequences (Chaudhari & Hahn, 1983).