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培养的人皮肤成纤维细胞中细胞视黄酸结合蛋白的证实

Demonstration of cellular retinoic acid binding protein in cultured human skin fibroblasts.

作者信息

Oikarinen A I, Oikarinen H, Uitto J

出版信息

Br J Dermatol. 1985 Nov;113(5):529-35. doi: 10.1111/j.1365-2133.1985.tb02376.x.

Abstract

Previous studies have indicated that retinoids, such as all-trans-retinoic acid and 13-cis-retinoic acid, can modulate connective tissue metabolism in human skin fibroblast cultures. Such effects could be mediated through binding of these retinoids to specific cellular binding proteins. In the present study we have demonstrated cellular retinoic acid binding protein using both whole cell and cytosol binding assays with [3H]all-trans-retinoic acid or [3H]13-cis-retinoic acid as the ligand. Specific binding of [3H]all-trans-retinoic acid could be demonstrated by both techniques and the binding could be displaced by unlabelled all-trans-retinoic acid and 13-cis-retinoic acid, but not by retinol or RO-10-9359 (etretinate) in a 100-fold excess. Gel filtration chromatography of the cytosol proteins after incubation with [3H]all-trans-retinoic acid demonstrated that the specific binding protein had an apparent molecular weight of approximately 15 000 daltons. Thus, the cellular retinoic acid binding protein demonstrated in human skin fibroblasts may mediate the effects of the retinoids on connective tissue metabolism in these cells.

摘要

先前的研究表明,类视黄醇,如全反式维甲酸和13 - 顺式维甲酸,可调节人皮肤成纤维细胞培养物中的结缔组织代谢。这些作用可能是通过这些类视黄醇与特定细胞结合蛋白的结合来介导的。在本研究中,我们使用以[3H]全反式维甲酸或[3H]13 - 顺式维甲酸作为配体的全细胞和胞质溶胶结合试验,证明了细胞视黄酸结合蛋白。两种技术都能证明[3H]全反式维甲酸的特异性结合,且这种结合可被未标记的全反式维甲酸和13 - 顺式维甲酸取代,但不能被过量100倍的视黄醇或RO - 10 - 9359(依曲替酯)取代。用[3H]全反式维甲酸孵育后的胞质溶胶蛋白进行凝胶过滤层析表明,特异性结合蛋白的表观分子量约为15000道尔顿。因此,在人皮肤成纤维细胞中证明的细胞视黄酸结合蛋白可能介导类视黄醇对这些细胞中结缔组织代谢的作用。

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