CHINESE-HAMSTER METAPHASE CHROMOSOMES ISOLATED UNDER PHYSIOLOGICAL CONDITIONS - A PARTIAL CHARACTERIZATION OF ASSOCIATED NON-HISTONE PROTEINS AND PROTEIN CORES
CHINESE-HAMSTER METAPHASE CHROMOSOMES ISOLATED UNDER PHYSIOLOGICAL CONDITIONS - A PARTIAL CHARACTERIZATION OF ASSOCIATED NON-HISTONE PROTEINS AND PROTEIN CORES
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DOI：
10.1016/0014-4827(83)90435-4
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发表时间：
1983-01-01
影响因子：
3.7
通讯作者：
JEPPESEN, P
中科院分区：
文献类型：
--
作者：
GOODERHAM, K;JEPPESEN, P
本文报道了一种从中国仓鼠中期染色体中分离得到的非组蛋白核心。这种核心结构保持了中期染色体的整体形态，即使在去除了所有的组蛋白，以及许多非组蛋白蛋白和大部分DNA之后。开发了一种用于分离中期染色体的新方法，该方法避免了使用高pH缓冲液和己二醇，并且消除了先前采用的众多离心和重悬步骤。通过2 M NaCl提取和DNase I消化从这些更温和的准生理条件下分离的中期染色体制备的染色体核心显示含有相对简单的非组蛋白的子集。一维SDS[十二烷基硫酸钠]-聚丙烯酰胺凝胶电泳显示了两组主要的多肽，分别具有MW 48，000 - 52，000和65，000 - 72，000 D [道尔顿]，其迁移率与核孔复合物-纤层多肽和微管蛋白相似。通过二维凝胶电泳和肽图谱的更详细的分析未能检测到这些蛋白质。核心的52，000 D多肽组分暂时被鉴定为中间丝蛋白波形蛋白。讨论了染色体核心的体内意义。
A non-histone protein core was described which could be isolated from Chinese hamster metaphase chromosomes. This core structure maintained the overall morphology of the metaphase chromosome even after removal of all of the histones, together with many of the non-histone proteins and the bulk of the DNA. A novel procedure was developed for the isolation of metaphase chromosomes which avoids the use of high pH buffers and hexylene glycol, as well as eliminating the numerous centrifugation and resuspension steps previously employed. Chromosome cores prepared by 2 M NaCl extraction and DNase I digestion from metaphase chromosomes isolated under these more gentle, quasi-physiological conditions, are shown to contain a relatively simple subset of non-histone proteins. One-dimensional SDS[sodium dodecyl sulfate]-polyacrylamide gel electrophoresis shows 2 major groups of polypeptides having MW 48,000-52,000 and 65,000-72,000 D [dalton], respectively, with similarities in mobilities to the nuclear pore complex-lamina polypeptides and tubulins. More detailed analysis by 2-dimensional gel electrophoresis and peptide mapping has failed to detect these proteins. A 52,000 D polypeptide component of the core is tentatively identified as the intermediate filament protein vimentin. The in vivo significance of chromosome cores is discussed.