Author:
Date:
2015-02-20
[Abstract] This protocol describes a robust technique for the measurement of ZMYND11-DNA interaction by a label-free Biolayer Interferometry (BLI). ZMYND11 is a novel histone reader protein that specifically recognizes H3.3K36me3 via its tandem Bromodomain, zinc-finger and PWWP domain (BP). ZMYND11 links the histone-variant-mediated transcription elongation control to tumour suppression and may therefore represent a novel class of drug targets. Like other PWWP domains, ZMYND11 PWWP domain shows highly positively charged surface and interacts with DNA. Previously reported methods include NMR, FP or EMSA. Biolayer interferometry (BLI) is an emerging technology for analyzing all kinds of biomolecular interactions, such as protein-protein and protein-DNA binding. BLI allows for the real time monitoring ...
[摘要] 该协议描述了用于通过无标记生物分子干涉测量法(BLI)测量ZMYND11-DNA相互作用的鲁棒技术。 ZMYND11是一种新型组蛋白阅读器蛋白,通过其串联溴结构域,锌指和PWWP结构域(BP)特异性识别H3.3K36me3。 ZMYND11将组蛋白变体介导的转录伸长控制与肿瘤抑制联系起来,因此可能代表一种新型的药物靶标。像其他PWWP域一样,ZMYND11 PWWP域显示高度带正电荷的表面,并与DNA相互作用。先前报道的方法包括NMR,FP或EMSA。生物层干涉法(BLI)是用于分析各种生物分子相互作用(例如蛋白质 - 蛋白质和蛋白质-DNA结合)的新兴技术。 BLI允许实时监测生物分子之间的相互作用,而不需要具有酶,荧光或放射性标记的试剂。该技术基于当材料结合到光纤的尖端时从光纤的表面反射的光的干涉图案的变化。该技术代表了诸如表面等离子体共振等技术的替代方法,提供了一种简单的平台,其能够在不使用流动池的情况下实现生物分子相互作用的无标记监测。无标记的生物传感器方法提供关于结合,动力学,浓度和相互作用的亲和力的信息。
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