Guy Hill, Ji-Chun Yang, Laura Easton, Rachel Cerdan, Stephen McLaughlin, Katherine Stott, Andrew Travers, David Neuhaus
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引用次数: 0
摘要
在应用溶液核磁共振确定复合物的结构时,界面处原子信号的增宽往往是一个限制因素。造成此类问题的常见原因包括自由态和结合态之间的交换以及相对于自由成分而言复合物分子量的增加,但当界面内的构象动态以化学位移时间尺度上的中间速率发生时,另一个更难处理的原因就会出现。在这项研究中,我们展示了如何通过对蛋白质 HMG-D 进行精心选择的突变来挽救这种情况,从而能够对其与模仿天然 DNA 弯曲而设计的 dA2 隆起 DNA 配体的复合物进行高分辨率结构测定,进而揭示复合物的新空间组织。
A Single Interfacial Point Mutation Rescues Solution Structure Determination of the Complex of HMG-D with a DNA Bulge.
Broadening of signals from atoms at interfaces can often be a limiting factor in applying solution NMR to the structure determination of complexes. Common contributors to such problems include exchange between free and bound states and the increased molecular weight of complexes relative to the free components, but another cause that can be more difficult to deal with occurs when conformational dynamics within the interface takes place at an intermediate rate on the chemical shift timescale. In this work we show how a carefully chosen mutation in the protein HMG-D rescued such a situation, making possible high-resolution structure determination of its complex with a dA2 bulge DNA ligand designed to mimic a natural DNA bend, and thereby revealing a new spatial organization of the complex.
期刊介绍:
ChemBioChem (Impact Factor 2018: 2.641) publishes important breakthroughs across all areas at the interface of chemistry and biology, including the fields of chemical biology, bioorganic chemistry, bioinorganic chemistry, synthetic biology, biocatalysis, bionanotechnology, and biomaterials. It is published on behalf of Chemistry Europe, an association of 16 European chemical societies, and supported by the Asian Chemical Editorial Society (ACES).