BERBERINE SULFATE BINDING TO MAST-CELL POLYANIONS - CYTOFLUOROMETRIC METHOD FOR QUANTITATION OF HEPARIN
BERBERINE SULFATE BINDING TO MAST-CELL POLYANIONS - CYTOFLUOROMETRIC METHOD FOR QUANTITATION OF HEPARIN
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DOI：
10.1007/bf00492678
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发表时间：
1974-01-01
期刊：
影响因子：
--
通讯作者：
ENERBACK, L
中科院分区：
文献类型：
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作者：
ENERBACK, L
本文报道了荧光阳离子染料硫酸黄连素与某些组织阴离子结合的细胞荧光研究。对腹膜细胞和含有肝素或DNA的模型进行测量。发现该染料适用于肝素的细胞荧光定量。在pH 4时，当染料明显选择性地与含磷酸盐和硫酸盐的聚阴离子结合时，建立了染色平衡。与DNA的结合比与肝素的结合更耐盐提取。阻断实验表明，大多数的肥大细胞的硫酸肝素可用于染料结合，而核DNA的磷酸基团在很大程度上被碱性蛋白质基团所阻断。由与核酸结合产生的荧光估计为平均肥大细胞的荧光强度的2%或更小。模型实验表明，肝素的量和荧光强度之间的线性关系。荧光光谱显示，染料与肝素结合时的最大发射值略高于与DNA结合时的最大发射值，表明染料以相关分子形式与肝素结合。这项研究还揭示了一些有趣的，但尚未解释的差异，在所产生的荧光性质时，染料结合到不同的聚阴离子。肝素结合的硫酸黄连素表现出快速的指数衰减后，连续照明，无论是在模型和肥大细胞，而DNA结合的染料表现出缓慢增加的荧光强度在连续照明。
A cytofluorometric study on the binding of the fluorescent cationic dye Berberine sulphate to some tissue anions is reported. Measurements were performed on peritoneal cells and on models containing heparin or DNA. The dye was found to be suitable for cytofluorometric quantitation of heparin. At pH 4 a staining equilibrium was established when the dye apparently binds selectively to phosphate and sulphate containing polyanions. Binding to DNA was more resistant to salt extraction than the binding to heparin. Blocking experiments demonstrated that most of the heparin sulphate of mast cells was available for dye binding while the phosphate groups of nuclear DNA were largely blocked by basic protein groups. The fluorescence resulting from binding to nucleic acids was estimated to be 2% or less of the fluorescence intensity of an average mast cell. Model experiments demonstrated a linear relationship between the amount of heparin and the intensity of fluorescence. Fluorescence spectroscopy showed that the dye had a slightly higher emission maximum when bound to heparin than to DNA indicating that the dye binds to heparin in an associated molecular form. The study also revealed some interesting, although yet unexplained differences in the properties of the resulting fluorescence when the dye was bound to different polyanions. Heparin-bound Berberine sulphate showed a rapid exponential fading upon continuous illumination, both in models and in mast cells, while the DNA bound dye showed a slow increase in fluorescence intensity during continuous illumination.