Two-dimensinal gel electrophoresis of rat liver nuclear washes, nuclear matrix, and hnRNA proteins.
Two-dimensinal gel electrophoresis of rat liver nuclear washes, nuclear matrix, and hnRNA proteins.
复制标题
DOI：
10.1083/jcb.86.1.135
复制
发表时间：
1980-07
期刊：
影响因子：
--
通讯作者：
Commings DE
中科院分区：
文献类型：
--
作者：
Peters KE;Commings DE
用双向凝胶电泳法检测了大鼠肝细胞浆、核洗液、基质、膜、异质性核糖核酸蛋白和染色质的蛋白质。凝胶中包含了精心挑选的六种常见蛋白质标准的分子量和等电点，使我们能够清楚地跟踪核提取过程中特定蛋白质的分布。在核洗涤和染色质中，我们观察到五类蛋白质：(A)专有细胞质蛋白，存在于第一次生理盐水-EDTA洗涤中，但在随后的洗涤中迅速消失；(B)普遍存在的75,000、68,000、57,000和43,000摩尔重量的蛋白质，后者是肌动蛋白，存在于细胞质、所有核洗涤和最终染色质颗粒中；(C)94,000、25,000和20,500摩尔重量的蛋白质；(D)核洗涤和最终染色质中存在的蛋白质，主要来自核基质中62,000、55,000、54,000和48,000摩尔重量的物种；和(E)只存在于最终染色质中的两种68000molwt的蛋白质。核基质的一维凝胶电泳法显示，主要的65,000-75,000摩尔的wt蛋白是非常不均匀的，包含一个主要的酸性基团，一个中间基团和一个碱性基团。单一的68,000摩尔重量的多肽构成了膜-板层部分的大部分，这与免疫学研究一致，表明在细胞核的外围存在与异染色质相关的一组不同的基质蛋白。肌动蛋白是第二种主要的核膜-板层蛋白。两个分子量分别为36,000和34,000的多肽占hnRNP的60%。未经洗涤的细胞核中约80%的非组蛋白染色体蛋白(NHP)由核基质和hnRNPs贡献，染色质NHP的模式基本相同。NHP是一组不同的DNA结合蛋白质的概念是不必要的限制。许多是从核基质或hnRNP颗粒中衍生出来的，它们共享不同的细胞内隔室的程度不同。
The proteins of rat liver cytoplasm, nuclear washes, matrix, membrane, heterogeneous nuclear (hn)RNA proteins and chromatin were examined by two-dimensional gel electrophoresis. The inclusion in the gels of six common protein standards of carefully selected molecular weight and isoelectric point allowed us to clearly follow the distribution of specific proteins during nuclear extraction. In the nuclear washes and chromatin, we observed five classes of proteins: (a) Exclusively cytoplasmic proteins, present in the first saline-EDTA wash but rapidly disappearing from subsequent washes; (b) ubiquitous proteins of 75,000, 68,000, 57,000, and 43,000 mol wt, the latter being actin, found in the cytoplasm, all nuclear washes and the final chromatin pellet; (c) proteins of 94,000, 25,000, and 20,500 mol wt specific to the nuclear washes; (d) proteins present in the nuclear washes and final chromatin, represented by species at 62,000, 55,000, 54,000, and 48,000 mol wt, primarily derived from the nuclear matrix; and (e) two proteins of 68,000 mol wt present only in the final chromatin. The major 65,000- 75,000-mol wt proteins seen by one-dimensional gel electrophoresis of nuclear matrix were very heterogeneous and contained a major acidic, an intermediate, and a basic group. A single 68,000-mol wt polypeptide constituted the majority of the membrane-lamina fraction, consistent with immunological studies indicating that a distinct subset of matrix proteins occurs, associated with heterochromatin, at the periphery of the nucleus. Actin was the second major nuclear membrane-lamina protein. Two polypeptides at 36,000 and 34,000 mol wt constituted 60% of the hnRNP. Approximately 80% of the mass of the nonhistone chromosomal proteins (NHP) from unwashed nuclei is contributed by nuclear matrix and hnRNPs, and essentially the same patterns were seen with chromatin NHP. The concept of NHP being a distinct set of DNA- bound proteins is unnecessarily limiting. Many are derived from the nuclear matrix or hnRNp particles and vary in the degree to which they share different intracellular compartments.