与26,26,26,27,27,27-六氟-1,25-二羟基维生素D3相关的维生素D3受体的DNA结合特性

DNA binding property of vitamin D3 receptors associated with 26,26,26,27,27,27-hexafluoro-1,25-dihydroxyvitamin D3.

作者信息

Inaba M, Okuno S, Inoue A, Nishizawa Y, Morii H, DeLuca H F

机构信息

Second Department of Internal Medicine, Osaka City University Medical School, Japan.

出版信息

Arch Biochem Biophys. 1989 Jan;268(1):35-9. doi: 10.1016/0003-9861(89)90562-6.

DOI:10.1016/0003-9861(89)90562-6

PMID:2463789

Abstract

Using [3H]-26,26,26,27,27,27-hexafluoro-1,25-dihydroxyvitamin D3 (F6-1,25-(OH)2D3), we have examined its ability to bind to the 1,25-(OH)2D3 receptor, and the ability of the resulting complex to bind DNA. The binding sites for [3H]F6-1,25-(OH)2D3 in the chick intestinal receptor represented a limited number of saturable sites for which 1,25-(OH)2D3 competes. 1,25-Dihydroxyvitamin D3 is three times more active than F6-1,25-(OH)2D3 in displacing [3H]F6-1,25-(OH)2D3. By affinity chromatography using DNA-Sephadex, the [3H]F6-1,25-(OH)2D3 receptor complex eluted from the column in a single peak at 0.14 M KCl, while [3H]-1,25-(OH)2D3 receptor complex eluted at 0.13 M KCl. These results indicate that F6-1,25-(OH)2D3 and 1,25-(OH)2D3 recognize the same binding site of the receptor and that the F6-1,25-(OH)2D3 receptor complex binds DNA more tightly than the 1,25-(OH)2D3 receptor complex. We suggest that the higher binding affinity for DNA may contribute to the greater biological activity of F6-1,25-(OH)2D3.

摘要

我们使用[3H]-26,26,26,27,27,27-六氟-1,25-二羟基维生素D3（F6-1,25-(OH)2D3），检测了其与1,25-(OH)2D3受体结合的能力，以及所得复合物与DNA结合的能力。雏鸡肠道受体中[3H]F6-1,25-(OH)2D3的结合位点代表了有限数量的可饱和位点，1,25-(OH)2D3可与之竞争。在取代[3H]F6-1,25-(OH)2D3方面，1,25-二羟基维生素D3的活性是F6-1,25-(OH)2D3的三倍。通过使用DNA-葡聚糖凝胶的亲和层析，[3H]F6-1,25-(OH)2D3受体复合物在0.14M KCl浓度下以单峰形式从柱上洗脱，而[3H]-1,25-(OH)2D3受体复合物在0.13M KCl浓度下洗脱。这些结果表明，F6-1,25-(OH)2D3和1,25-(OH)2D3识别受体的相同结合位点，并且F6-1,25-(OH)2D3受体复合物比1,25-(OH)2D3受体复合物更紧密地结合DNA。我们认为，对DNA更高的结合亲和力可能有助于F6-1,25-(OH)2D3具有更强的生物学活性。