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Development of reducing western blot to detect rodox conformers of hHSF1
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF EMERGENCY MEDICINE
- Issue:
- 5
- DOI:
- 10.3760/j.issn:1671-0282.2003.05.006
- Key Word:
- 热休克转录因子1;Western blot;氧化还原构像
Abstract： 目的分辨、检测人类热休克转录因子1(hHSF1)的氧化还原构像,以了解构像的改变对其功能的影响.方法建立了一种新型的还原型Western blot检测法,使已形成稳定屏障式抗原核心,双硫键交联的氧化型人类热休克转录因子1(ox-hHSF1)还原,恢复其免疫检测活性.结果无论在变性聚丙烯酰胺凝胶电泳或非变性聚丙烯酰胺凝胶电泳的还原型Western blot中都可检测出一个在常规不能检测出的、移动快速、结构致密、分子内双硫键交联的ox-hHSF1构像.ox-hHSF1的出现,使hHSF1在体外不能被热诱导激活形成三体,失去其与DNA结合的功能.结论还原型Western blot的建立,能更进一步帮助了解氧化还原状态的改变怎样影响包括hHSF1在内的转录因子结构和功能.
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