Analysis of sugar-binding sites in mammalian cell nuclei by quantitative flow microfluorometry.
Analysis of sugar-binding sites in mammalian cell nuclei by quantitative flow microfluorometry.
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通过定量流式显微荧光法分析哺乳动物细胞核中的糖结合位点。
DOI：
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发表时间：
1986
影响因子：
11.1
通讯作者：
M. Monsigny
中科院分区：
文献类型：
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作者：
A. Seve;J. Hubert;D. Bouvier;C. Bourgeois;P. Midoux;A. Roche;M. Monsigny
新糖蛋白（偶联到糖和荧光素的牛血清白蛋白）结合的定量流动显微荧光测定法证明了糖结合位点的存在（即，凝集素样分子）。通过细胞裂解分离的细胞核和通过去核获得的透性化的核质体获得的非常相似的标记强度加强了结合位点由实际的细胞核结构而不是由细胞质或膜衍生的污染物承担的想法。使用两种核分离程序，用作标记物的新糖蛋白（含有相似数量的糖残基）可以类似地分类。荧光显微镜进一步表明，在这两个核制剂，neoglycoprotein结合位点与核仁以及与核质核糖核蛋白元素。从指数增长的细胞的细胞核绑定更大量的新糖蛋白比接触抑制细胞的细胞核。
Quantitative flow microfluorometry of neoglycoprotein (bovine serum albumin coupled to sugar and to fluorescein) binding demonstrated the existence of sugar-binding sites (i.e., lectin-like molecules) in isolated BHK cell nuclei. The very similar labeling intensities obtained with nuclei isolated by cell lysis and with permeabilized karyoplasts obtained by enucleation strengthened the idea that the binding sites are borne by actual nuclear structures and not by cytoplasmic or membrane-derived contaminants. With both nuclei-isolation procedures, neoglycoproteins (containing similar numbers of sugar residues) used as markers can be similarly classified. Fluorescence microscopy further indicated that in both nuclear preparations, the neoglycoprotein binding sites were associated with the nucleoli as well as with nucleoplasmic ribonucleoprotein elements. Nuclei from exponentially growing cells bound much greater amounts of neoglycoprotein than did nuclei from contact-inhibited cells.