Bourgeois C A, Monsigny M
Départment de Biochimie des Glycoconjugués, Centre de Biophysique Moléculaire, Orléans, France.
Biol Cell. 1990;69(2):119-26. doi: 10.1111/j.1768-322x.1990.tb00007.x.
Using gold labelled neoglycoproteins containing either alpha-D-glucose, N-acetyl-beta-D-glucosamine, alpha-D-mannose, 6-phospho-alpha-D-mannose, and alpha-L-fucose (BSA), we investigated their intranuclear binding sites in the TG human cell line. Although gold-labelled BSA did not give any noticeable labelling, the presence of 1% free BSA in the medium containing the gold labelled neoglycoproteins was revealed to be a key factor of the labelling. During interphase in the presence of free BSA most of the labelling was detected in the nucleoplasm. The border of the condensed chromatin, known to be the site of hnRNA synthesis as well as the interchromatin areas enriched in RNPs were labelled. Condensed chromatin also contained binding-sites. The nucleolus was seen to present low labelling in comparison with the labelling observed over the nucleoplasm. These nucleolar binding sites were located both in the dense fibrillar and granular components. No labelling could be detected over the fibrillar centers which are very conspicuous in this cell line. During mitosis sugar-binding sites were observed over the chromosomes. Data reported here show for the first time that lectin-like proteins and chromatin components are colocalized both during interphase and mitosis. In addition, within the nucleolus the presence of sugar-binding proteins was seen to be restricted to the dense fibrillar and granular components.
我们使用了含有α-D-葡萄糖、N-乙酰-β-D-葡萄糖胺、α-D-甘露糖、6-磷酸-α-D-甘露糖和α-L-岩藻糖(牛血清白蛋白)的金标记新糖蛋白,研究了它们在TG人细胞系中的核内结合位点。尽管金标记的牛血清白蛋白没有给出任何明显的标记,但发现在含有金标记新糖蛋白的培养基中存在1%的游离牛血清白蛋白是标记的关键因素。在有游离牛血清白蛋白存在的间期,大部分标记出现在核质中。已知是hnRNA合成位点的浓缩染色质边界以及富含核糖核蛋白的染色质间区域被标记。浓缩染色质也含有结合位点。与在核质上观察到的标记相比,核仁的标记较低。这些核仁结合位点位于致密纤维成分和颗粒成分中。在该细胞系中非常明显的纤维中心未检测到标记。在有丝分裂期间,在染色体上观察到糖结合位点。此处报道的数据首次表明,类凝集素蛋白和染色质成分在间期和有丝分裂期间共定位。此外,在核仁内,糖结合蛋白的存在仅限于致密纤维成分和颗粒成分。