Romani M, Vidali G, Tahourdin C S, Bustin M
J Biol Chem. 1980 Jan 25;255(2):468-74.
This manuscript describes the use of a solid phase radioimmunoassay for serological analysis of chromosomal components. The applicability of this assay for various studies on nonhistone chromosomal proteins, histones, and chromatin subunits is illustrated. By this technique it is possible to detect and quantify nuclear antigens in the nanogram range. The assay has all the inherent sensitivity and precision of radioimmunoassays and, as such, introduces a new, convenient method for serological analyses of chromosomal components. The results presented reconfirm the serological similarity among the HMG (high mobility group) proteins derived from various sources. The amount of HMG proteins present in mononucleosomes purified from calf thymus is similar to that present in mononucleosomes purified from HeLa cells, suggesting that various tissues contain similar amounts of these proteins. Per nucleosome, dinucleosomes and trinucleosomes contain as much HMG-1 protein as mononucleosomes, suggesting that the protein is not exclusively associated with those regions of DNA which have been solubilized by micrococcal nuclease. Part of the antigenic determinants present in HMG-1 forming a complex in the nucleosomal conformation do not interact with antibodies.
本手稿描述了一种用于染色体成分血清学分析的固相放射免疫测定法。阐述了该测定法在各种非组蛋白染色体蛋白、组蛋白和染色质亚基研究中的适用性。通过这种技术,可以检测和定量纳克级的核抗原。该测定法具有放射免疫测定法固有的所有灵敏度和精密度,因此为染色体成分的血清学分析引入了一种新的、便捷的方法。所呈现的结果再次证实了源自各种来源的HMG(高迁移率族)蛋白之间的血清学相似性。从小牛胸腺纯化的单核小体中存在的HMG蛋白量与从HeLa细胞纯化的单核小体中存在的量相似,这表明各种组织中这些蛋白的含量相似。每个核小体、双核小体和三核小体所含的HMG-1蛋白与单核小体一样多,这表明该蛋白并非仅与那些已被微球菌核酸酶溶解的DNA区域相关联。以核小体构象形成复合物的HMG-1中存在的部分抗原决定簇不与抗体相互作用。