Schulte T W, Akinaga S, Soga S, Sullivan W, Stensgard B, Toft D, Neckers L M
Medicine Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Cell Stress Chaperones. 1998 Jun;3(2):100-8. doi: 10.1379/1466-1268(1998)003<0100:arbttn>2.3.co;2.
The molecular chaperone Hsp90 plays an essential role in the folding and function of important cellular proteins including steroid hormone receptors, protein kinases and proteins controlling the cell cycle and apoptosis. A 15 A deep pocket region in the N-terminal domain of Hsp90 serves as an ATP/ADP-binding site and has also been shown to bind geldanamycin, the only specific inhibitor of Hsp90 function described to date. We now show that radicicol, a macrocyclic antifungal structurally unrelated to geldanamycin, also specifically binds to Hsp90. Moreover, radicicol competes with geldanamycin for binding to the N-terminal domain of the chaperone, expressed either by in vitro translation or as a purified protein, suggesting that radicicol shares the geldanamycin binding site. Radicicol, as does geldanamycin, also inhibits the binding of the accessory protein p23 to Hsp90, and interferes with assembly of the mature progesterone receptor complex. Radicicol does not deplete cells of Hsp90, but rather increases synthesis as well as the steady-state level of this protein, similar to a stress response. Finally, radicicol depletes SKBR3 cells of p185erbB2, Raf-1 and mutant p53, similar to geldanamycin. Radicicol thus represents a structurally unique antibiotic, and the first non-benzoquinone ansamycin, capable of binding to Hsp90 and interfering with its function.
分子伴侣Hsp90在重要细胞蛋白的折叠和功能中发挥着关键作用,这些蛋白包括类固醇激素受体、蛋白激酶以及控制细胞周期和凋亡的蛋白。Hsp90 N端结构域中一个15埃深的口袋区域作为ATP/ADP结合位点,并且已被证明还能结合格尔德霉素,这是迄今为止所描述的Hsp90功能的唯一特异性抑制剂。我们现在表明,雷迪西醇,一种在结构上与格尔德霉素无关的大环抗真菌剂,也能特异性地结合Hsp90。此外,雷迪西醇与格尔德霉素竞争结合伴侣蛋白的N端结构域,该结构域通过体外翻译或作为纯化蛋白表达,这表明雷迪西醇与格尔德霉素共享结合位点。与格尔德霉素一样,雷迪西醇也抑制辅助蛋白p23与Hsp90的结合,并干扰成熟孕酮受体复合物的组装。雷迪西醇不会使细胞中的Hsp90耗尽,而是增加这种蛋白的合成以及稳态水平,类似于应激反应。最后,雷迪西醇使SKBR3细胞中的p185erbB2、Raf-1和突变型p53耗尽,类似于格尔德霉素。因此,雷迪西醇代表了一种结构独特的抗生素,也是第一种能够结合Hsp90并干扰其功能的非苯醌安莎霉素。